Machine harvestable iceberg lettuce

ABSTRACT

Certain aspects of the present disclosure relate to upright heading iceberg lettuce plants including a space between the base of the head and the top of the ground, wherein the space includes an increased length of core outside of the processing material of the head (i.e., external stem). The increased external stem lengths of the upright heading iceberg lettuce plants of the present disclosure make them suitable for machine harvesting. Other aspects of the present disclosure relate to methods of generating and selecting upright heading iceberg lettuce plants. New upright heading iceberg lettuce varieties designated ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, and ‘E01E.70168’ Lot B are described.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 62/609,261, filed Dec. 21, 2017, and U.S. ProvisionalPatent Application No. 62/609,269, filed Dec. 21, 2017, both of whichare hereby incorporated by reference in their entireties.

FIELD OF THE INVENTION

The present disclosure relates to the field of plant breeding. Inparticular, the present disclosure relates to new and distinctivemachine harvestable iceberg lettuce plants (Lactuca sativa).

BACKGROUND

Cultivated forms of lettuce belong to the highly polymorphic speciesLactuca sativa that is grown for its edible head and leaves. As a crop,lettuce is grown commercially wherever environmental conditions permitthe production of an economically viable yield. For planting purposes,the lettuce season is typically divided into three categories (i.e.,early, mid, and late), with coastal areas planting from January toAugust, and desert regions planting from August to December. Freshlettuce is consumed nearly exclusively as fresh, raw product andoccasionally as a cooked vegetable.

Lactuca sativa is in the Cichorieae tribe of the Asteraceae (Compositae)family. Lettuce is related to chicory, sunflower, aster, dandelion,artichoke, and chrysanthemum. Sativa is one of about 300 species in thegenus Lactuca. There are seven different morphological types of lettuce.The crisphead group includes the iceberg and batavian types. Iceberglettuce has a large, firm head with a crisp texture and a white orcreamy yellow interior. The batavian lettuce predates the iceberg typeand has a smaller and less firm head. The butterhead group has a small,soft head with an almost oily texture. The romaine, also known as coslettuce, has elongated upright leaves forming a loose, loaf-shaped headand the outer leaves are usually dark green. Leaf lettuce, also known ascutting lettuce, comes in many varieties, none of which form a head, andinclude the green oak leaf variety. Latin lettuce, also known asgrasse-type lettuce, looks like a cross between romaine and butterhead.Stem lettuce has long, narrow leaves and thick, edible stems. Oilseedlettuce is a type grown for its large seeds that are pressed to obtainoil.

While iceberg lettuce is favored by consumers for the crisp texture ofits leaves and its taste, iceberg heads are difficult to grow andprocess. For one, the growth of iceberg lettuce close to the ground, aswell as its round shape, means that the heads must be harvested by hand.Machines are unable to cut iceberg varieties at the correct level (i.e.on the stem and sufficiently close to the ground), and so a largeproportion of processing material is lost. More upright varieties, suchas romaine, can be easily machine harvested because most of theirprocessing material sits well above the ground. For another, the closelypacked and tightly wrapped leaves of iceberg varieties mean thatmoisture is trapped within the iceberg head. This high level of moisturemeans that iceberg varieties are more susceptible to pathogens than moreopen varieties such as romaine. Finally, the round shape of iceberg aswell as its tightly packed leaves mean that iceberg is primarily used inchopped salads. The iceberg head cannot be cleaned thoroughly enough,and the iceberg leaves are too deeply cupped to be used as individualleaves. In contrast, the open and upright stature of romaine allows easycleaning and separation of individual leaves.

Lettuce is an increasingly popular crop. As worldwide lettuceconsumption continues to increase, and the cost of labor continues torise, there is a need for a new type of lettuce that combines thequalities of iceberg lettuce with the ability to be machine harvested.In particular, there is a need for a new type of lettuce with an uprightstature and crisp-textured leaves that in addition is stable, highyielding, and agronomically sound.

SUMMARY OF THE INVENTION

In order to meet these needs, the present disclosure is directed toupright heading iceberg lettuce. In certain aspects, the presentdisclosure relates to an upright heading iceberg lettuce plant with aheight to diameter ratio greater than or equal to about 1.0, a height ofabout 1.5 to about 3 times a height of a standard iceberg lettuce plant,a crisp leaf texture, and a closed head. In some embodiments, theupright heading iceberg lettuce plant further includes one or morecharacteristics from the group of: a short core, a low level of internaltipburn, a low level of bottom rot, an increased number of leaves, afast fill rate of leaves in a head, and a space between a base of a headand a top of a ground. In some embodiments, the space between the baseof the head and the top of the ground is achieved by an increased lengthof core or an increased number of frame leaves outside of the processingmaterial of a head.

Certain aspects of the present disclosure relate to an iceberg lettuceseed, wherein the seed produces an iceberg lettuce plant with an uprightstature phenotype. In some embodiments, the upright stature phenotypeincludes a height to diameter ratio greater than or equal to about 1.0,a height of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head. In someembodiments, the iceberg lettuce plant further includes one or morecharacteristics from the group of: a short core, a low level of internaltipburn, a low level of bottom rot, an increased number of leaves, afast fill rate of leaves in a head, and a space between a base of a headand a top of a ground.

Certain aspects of the present disclosure relate to an iceberg lettuceseed containing an upright stature allele at locus A, wherein the seedproduces an iceberg lettuce plant with an upright stature phenotype. Insome embodiments, the upright stature phenotype includes a height todiameter ratio greater than or equal to about 1.0, a height of about 1.5to about 3 times a height of a standard iceberg lettuce plant, a crispleaf texture, and a closed head. In some embodiments, the iceberglettuce plant further includes one or more characteristics from thegroup of: a short core, a low level of internal tipburn, a low level ofbottom rot, an increased number of leaves, a fast fill rate of leaves ina head, and a space between a base of a head and a top of a ground.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of a first iceberg lettuce plant with asecond iceberg lettuce plant, wherein the first iceberg lettuce plantproduces an upright heading iceberg lettuce plant with a height todiameter ratio greater than or equal to about 1.0, about 1.5 to about 3times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head, the second iceberg lettuce plant produces anupright heading iceberg lettuce plant with a height to diameter ratiogreater than or equal to about 1.0, about 1.5 to about 3 times a heightof a standard iceberg lettuce plant, a crisp leaf texture, and a closedhead, and wherein the iceberg lettuce plant produced from the cross hasa height to diameter ratio greater than or equal to about 1.0, a heightof about 1.5 to about 3 times a height of a standard iceberg lettuceplant, a crisp leaf texture, and a closed head. In one embodiment, thefirst and second iceberg lettuce plants are ‘E01E.70111’ Lot A lettuceplants, a sample of ‘E01E.70111’ Lot A lettuce seed having beendeposited under NCIMB Accession Number 42957. In another embodiment, thefirst and second iceberg lettuce plants are ‘E01E.70111’ Lot B lettuceplants, a sample of ‘E01E.70111’ Lot B lettuce seed having beendeposited under NCIMB Accession Number 42962. In one embodiment, thefirst and second iceberg lettuce plants are ‘E01E.70168’ Lot A lettuceplants, a sample of ‘E01E.70168’ Lot A lettuce seed having beendeposited under NCIMB Accession Number 42958. In another embodiment, thefirst and second iceberg lettuce plants are ‘E01E.70168’ Lot B lettuceplants, a sample of ‘E01E.70168’ Lot B lettuce seed having beendeposited under NCIMB Accession Number 42963. In still anotherembodiment, the first iceberg lettuce plant is an ‘E01E.70111’ Lot Alettuce plant, a sample of ‘E01E.70111’ Lot A lettuce seed having beendeposited under NCIMB Accession Number 42957. In a further embodiment,the second iceberg lettuce plant is an ‘E01E.70168’ Lot A lettuce plant,a sample of ‘E01E.70168’ Lot A lettuce seed having been deposited underNCIMB Accession Number 42958.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot A plant with a second‘E01E.70111’ Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42957, wherein theiceberg lettuce seed produced from the cross produces an iceberg lettucewith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot B plant with a second‘E01E.70111’ Lot B plant, a sample of ‘E01E.70111’ Lot B lettuce seedhaving been deposited under NCIMB Accession Number 42962, wherein theiceberg lettuce seed produced from the cross produces an iceberg lettucewith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70168’ Lot A plant with a second‘E01E.70168’ Lot A plant, a sample of ‘E01E.70168’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42958, wherein theiceberg lettuce seed produced from the cross produces an iceberg lettucewith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70168’ Lot B plant with a second‘E01E.70168’ Lot B plant, a sample of ‘E01E.70168’ Lot B lettuce seedhaving been deposited under NCIMB Accession Number 42963, wherein theiceberg lettuce seed produced from the cross produces an iceberg lettucewith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot A plant with a second‘E01E.70168’ Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42957, and a sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958, wherein the iceberg lettuce seed produced fromthe cross produces an iceberg lettuce with a height to diameter ratiogreater than or equal to about 1.0, a height of about 1.5 to about 3times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head.

In other aspects, the present disclosure provides an iceberg lettuceseed from the plants of any of the above embodiments. In anotherembodiment, the present disclosure is directed to iceberg lettuce plantsgrown from the seed of any of the above embodiments. In some embodimentsof any of the above embodiments, the iceberg lettuce plant has a heightto diameter ratio greater than or equal to about 1.0, a height of about1.5 to about 3 times a height of a standard iceberg lettuce plant, acrisp leaf texture, and a closed head. In some embodiments, the iceberglettuce plant further includes one or more characteristics from thegroup of: a crisp leaf texture, a short core, a low level of internaltipburn, a low level of bottom rot, an increased number of leaves, afast fill rate of leaves in a head, and a space between a base of a headand a top of a ground.

Lettuce plant parts include lettuce heads, lettuce leaves, parts oflettuce leaves, pollen, ovules, flowers, and the like. In anotherembodiment, the present disclosure is directed to a plant part of theplants of any of the above embodiments. In another embodiment, thepresent disclosure is further directed to heads, leaves, parts ofleaves, stems, roots, meristems, flowers, pollen, and ovules of theplants of any of the above embodiments. In another embodiment, thepresent disclosure is further directed to tissue culture of the plantsof any of the above embodiments, and to plants regenerated from thetissue culture, where the plant has all the morphological andphysiological characteristics of the plants of any of the aboveembodiments. In one embodiment, the plant regenerated from the tissueculture has all of the morphological and physiological characteristicsof an iceberg lettuce plant produced by growing seed designated as‘E01E.70111’ Lot A, representative sample of ‘E01E.70111’ Lot A lettuceseed having been deposited under NCIMB Accession Number 42957. Inanother embodiment, the plant regenerated from the tissue culture hasall of the morphological and physiological characteristics of an iceberglettuce plant produced by growing seed designated as ‘E01E.70111’ Lot B,representative sample of ‘E01E.70111’ Lot B lettuce seed having beendeposited under NCIMB Accession Number 42962. In one embodiment, theplant regenerated from the tissue culture has all of the morphologicaland physiological characteristics of an iceberg lettuce plant producedby growing seed designated as ‘E01E.70168’ Lot A, representative sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958. In another embodiment, the plant regeneratedfrom the tissue culture has all of the morphological and physiologicalcharacteristics of an iceberg lettuce plant produced by growing seeddesignated as ‘E01E.70168’ Lot B, representative sample of ‘E01E.70168’Lot B lettuce seed having been deposited under NCIMB Accession Number42963. In some embodiments, the present disclosure is directed to aplant part of any of the above embodiments of plants regenerated fromtissue culture. In some embodiments, the present disclosure is furtherdirected to heads, leaves, parts of leaves, stems, roots, meristems,flowers, pollen, and ovules of any of the above embodiments of plantsregenerated from tissue culture.

In still another embodiment, the present disclosure is further directedto packaging material containing plant parts of the plants of any of theabove embodiments. Such packaging material includes but is not limitedto boxes, plastic bags, etc. The plant parts of the plants of any of theabove embodiments may be combined with other plant parts of other plantvarieties.

In another embodiment, the present disclosure is directed to methods ofproducing an herbicide resistant lettuce plant by introducing a geneconferring herbicide resistance into a lettuce plant produced by growinglettuce seed of any of the above embodiments, where the gene is selectedfrom glyphosate, sulfonylurea, imidazolinone, dicamba, glufosinate,phenoxy proprionic acid, L-phosphinothricin, cyclohexone,cyclohexanedione, triazine, and benzonitrile. Certain embodiments arealso directed to herbicide resistant lettuce plants produced by suchmethods.

In another embodiment, the present disclosure is directed to methods ofproducing a pest or insect resistant lettuce plan by introducing a geneconferring pest or insect resistance into a lettuce plant produced bygrowing lettuce seed of any of the above embodiments, and to pest orinsect resistant lettuce plants produced by such methods. In certainembodiments, the gene conferring pest or insect resistance encodes aBacillus thuringiensis endotoxin.

In another embodiment, the present disclosure is directed to methods ofproducing a disease resistant lettuce plant by introducing a geneconferring disease resistance into a lettuce plant produced by growinglettuce seed of any of the above embodiments, and to disease resistantlettuce plants produced by such methods.

In another embodiment, the present disclosure is directed to methods ofproducing a lettuce plant with a value-added trait by introducing a geneconferring a value-added trait into a lettuce plant produced by growinglettuce seed of any of the above embodiments, where the gene encodes aprotein selected from a ferritin, a nitrate reductase, and a monellin.Certain embodiments are also directed to lettuce plants having avalue-added trait produced by such methods.

In another embodiment, the present disclosure provides for single geneconverted plants of any of the above embodiments. The single transferredgene may preferably be a dominant or recessive allele. Preferably, thesingle transferred gene will confer such traits as male sterility,herbicide resistance, insect or pest resistance, modified fatty acidmetabolism, modified carbohydrate metabolism, resistance for bacterial,fungal, or viral disease, male fertility, enhanced nutritional quality,and industrial usage. The single gene may be a naturally occurringlettuce gene.

In another embodiment, the present disclosure is directed to a method ofmaking iceberg lettuce seeds, by a) crossing the plant of any of theabove embodiments with another iceberg lettuce plant, and b) harvestingthe seed from the cross. In still another embodiment, the presentdisclosure is further directed to lettuce plants, lettuce parts from thelettuce plants, and seeds produced therefrom where the lettuce plant isisolated by the breeding method of the disclosure.

In yet another embodiment, the present disclosure is further directed toa method of selecting lettuce plants with an upright stature, by a)crossing a first lettuce plant with a height to diameter ratio greaterthan or equal to about 1.0, a height of about 1.5 to about 3 times aheight of a standard iceberg lettuce plant, a crisp leaf texture, and aclosed head with a second lettuce plant with a height to diameter ratiogreater than or equal to about 1.0, a height of about 1.5 to about 3times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head, b) selecting offspring from the cross thathave an improved set of characteristics including a height to diameterratio greater than or equal to about 1.5, a height greater than or equalto about 2 to about 3 times a height of a standard iceberg lettuceplant, a crisp leaf texture, and a more tightly closed head, c) selfingor sibbing the offspring, and d) repeating steps b) and c) for multiplegenerations to produce inbred lines with the improved set ofcharacteristics. In one embodiment, the characteristics used forselection in step b) further includes one or more characteristics fromthe group of: a short core, a low level of internal tipburn, a low levelof bottom rot, an increased number of leaves, a fast fill rate of leavesin a head, and a space between a base of a head and a top of a ground.In another embodiment, the space between the base of the head and thetop of the ground is achieved by an increased length of core or anincreased number of frame leaves outside of the processing material ofthe head. In still another embodiment, the present disclosure is furtherdirected to lettuce plants, lettuce parts from the lettuce plants, andseeds produced therefrom where the lettuce plant is isolated by thebreeding method of the disclosure.

In a further embodiment, the present disclosure relates to methods fordeveloping lettuce plants in a lettuce plant breeding program usingplant breeding techniques including recurrent selection, backcrossing,pedigree breeding, restriction fragment length polymorphism enhancedselection, genetic marker enhanced selection, and transformation. Seeds,lettuce plants, and parts thereof, produced by such breeding methods arealso part of the disclosure.

In order to meet these needs, the present disclosure is directed toupright heading iceberg lettuce. In certain aspects, the presentdisclosure relates to an iceberg lettuce (Lactuca sativa) plantcontaining a genetic determinant that leads to the iceberg lettuce planthaving an upright stature, wherein the genetic determinant is ascontained in a lettuce plant representative seed of which was depositedwith the NCIMB under accession numbers 42957, 42962, 42958, and 42963.In some embodiments, the genetic determinant is homozygously present. Insome embodiments, the genetic determinant is heterozygously present. Insome embodiments, the upright stature includes one or morecharacteristics from the group of: a ratio of plant height to diametergreater than about 1, a height of about 1.5 to about 3 times a height ofa standard iceberg lettuce plant, a crisp leaf texture, and a closedhead. In some embodiments, the lettuce further includes one or morecharacteristics from the group of: a short core, a low level of internaltipburn, a low level of bottom rot, an increased number of leaves, afast fill rate of leaves in a head, and a space between a base of a headand a top of a ground.

In certain aspects, the present disclosure relates to an iceberg lettuce(Lactuca sativa) plant containing a genetic determinant that leads tothe iceberg lettuce plant having an upright stature, wherein the uprightstature comprises one or more characteristics from the group of: a ratioof plant height to diameter greater than about 1, a height of about 1.5to about 3 times a height of a standard iceberg lettuce plant, a crispleaf texture, and a closed head, and wherein the genetic determinant isas contained in a lettuce plant representative seed of which wasdeposited with the NCIMB under accession numbers 42957, 42962, 42958,and 42963. In some embodiments, the genetic determinant is homozygouslypresent. In some embodiments, the genetic determinant is heterozygouslypresent. In some embodiments, the lettuce further includes one or morecharacteristics from the group of: a short core, a low level of internaltipburn, a low level of bottom rot, an increased number of leaves, afast fill rate of leaves in a head, and a space between a base of a headand a top of a ground.

Certain aspects of the present disclosure relate to the plants of any ofthe above embodiments, obtainable by crossing a first iceberg lettuceplant with a second iceberg lettuce plant, wherein at least one of thesaid plants comprises the genetic determinant as comprised in a lettuceplant representative seed of which was deposited with the NCIMB underaccession numbers 42957, 42962, 42958, and 42963, or a progeny plantthereof carrying the genetic determinant, and selecting, preferably inthe F₂ generation, for plants having an upright stature.

Certain aspects of the present disclosure relate to a seed from theplants of any of the above embodiments, wherein the seed contains thegenetic determinant as present in seeds of which a representative samplewas deposited with the NCIMB under accession numbers 42957, 42962,42958, or 42963.

Certain aspects of the present disclosure relate to a progeny plant fromplants of any of the above embodiments, wherein the plant contains thegenetic determinant as present in seeds of which a representative samplewas deposited with the NCIMB under accession numbers 42957, 42962,42958, or 42963. In some embodiments, the genetic determinant ishomozygously present. In some embodiments, the genetic determinant isheterozygously present.

Certain aspects of the present disclosure relate to a progeny plant fromthe seeds of any of the above embodiments, wherein the plant containsthe genetic determinant as present in seeds of which a representativesample was deposited with the NCIMB under accession numbers 42957,42962, 42958, or 42963. In some embodiments, the genetic determinant ishomozygously present. In some embodiments, the genetic determinant isheterozygously present.

Certain aspects of the present disclosure relate to a propagationmaterial derived from the plants of any of the above embodiments,wherein the plant contains the genetic determinant as present in seedsof which a representative sample was deposited with the NCIMB underaccession numbers 42957, 42962, 42958, or 42963.

Certain aspects of the present disclosure relate to a propagationmaterial derived from the seeds of any of the above embodiments, whereinthe plant contains the genetic determinant as present in seeds of whicha representative sample was deposited with the NCIMB under accessionnumbers 42957, 42962, 42958, or 42963.

Certain aspects of the present disclosure relate to a propagationmaterial capable of growing into a plant of any of the aboveembodiments. In some embodiments, the propagation material is amicrospore, pollen, ovary, ovule, embryo, embryo sac, egg cell, cutting,root, root tip, hypocotyl, cotyledon, stem, leaf, flower, anther, seed,meristem, protoplast, callus, or cell.

Certain aspects of the present disclosure relate to tissue culture fromthe plants of any of the above embodiments. In some embodiments, thetissue culture is from the propagation material capable of growing intoa plant of any of the above embodiments. In some embodiments, thepropagation material is a microspore, pollen, ovary, ovule, embryo,embryo sac, egg cell, cutting, root, root tip, hypocotyl, cotyledon,stem, leaf, flower, anther, seed, meristem, protoplast, callus, or cell.

Certain aspects of the present disclosure relate to a plant part fromthe plants of any of the above embodiments. In some embodiments, theplant part is a head, a leaf, or a portion thereof. In some embodiments,the plant part is a head.

In another embodiment, the present disclosure is directed to a method ofproducing an iceberg lettuce plant with an upright stature phenotype, bya) crossing a lettuce plant of any of the above embodiments with asecond iceberg lettuce plant, wherein said second iceberg lettuce plantis a non-upright stature iceberg lettuce plant, to produce progeny; b)using said progeny of a) in a back-crossing breeding program with theiceberg lettuce plant of any of the above embodiments as the recurrentparent for one or more generations, to produce a progeny iceberg lettuceplant with an upright stature phenotype. In some embodiments, theupright stature phenotype includes one or more characteristics from thegroup of: a ratio of plant height to diameter greater than about 1, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, a closed head, a short core, a lowlevel of internal tipburn, a low level of bottom rot, an increasednumber of leaves, a fast fill rate of leaves in a head, and a spacebetween a base of a head and a top of a ground.

In another embodiment, the present disclosure is directed to a method ofmaking iceberg lettuce seeds, by a) crossing the plant of any of theabove embodiments with another iceberg lettuce plant, and b) harvestingthe seed from the cross. In still another embodiment, the presentdisclosure is further directed to lettuce plants, lettuce parts from thelettuce plants, and seeds produced therefrom, wherein the lettuce plantis isolated by the breeding method of the disclosure.

In a further embodiment, the present disclosure relates to methods fordeveloping lettuce plants in a lettuce plant breeding program usingplant breeding techniques including recurrent selection, backcrossing,pedigree breeding, restriction fragment length polymorphism enhancedselection, genetic marker enhanced selection, and transformation. Insome embodiments, the present disclosure relates to plants that haveobtained the genetic constitution or gene complex of plants of any ofthe above embodiments that leads to the upright stature phenotype,either by crossing or by means of molecular biological techniques. Insome embodiments, the present disclosure relates to the progeny of theseplants that have maintained or acquired the trait of the upright statureof the invention. Seeds, lettuce plants, and parts thereof, produced bysuch breeding methods are also part of the disclosure.

In certain aspects, the present disclosure relates to an upright headingiceberg lettuce plant including a space between a base of a head and atop of a ground. In some embodiments, said space includes an increasedlength of core outside of a processing material of a head (i.e.,external stem). In some embodiments, the iceberg lettuce plant furtherincludes one or more characteristics selected from the group of: aheight to diameter ratio of about 1.3 to about 1.5, leaf strengthbetween about 300 to 400 grams, about 40 to 50% overlapping leavesinside the processing material of the head, a short stem inside of theprocessing material of the head (i.e., internal stem), leaves with about4.8 to 5.3° Brix, leaves with about 25 to 40 g/l total sugars, leaveswith about 4.5 to 5% dry matter weight, 5-10% processing damage per bagof processed plant material, and a range of about 35 to 42 days of shelflife of processed and packaged plant material.

Certain aspects of the present disclosure relate to an iceberg lettuceseed, wherein the seed produces an iceberg lettuce plant with an uprightstature phenotype. In some embodiments, the upright stature phenotypeincludes a space between a base of a head and a top of a ground. In someembodiments, said space includes an increased length of core outside ofa processing material of a head (i.e., external stem). In someembodiments, the upright stature phenotype further includes one or morecharacteristics selected from the group of: a height to diameter ratioof about 1.3 to about 1.5, leaf strength between about 300 to 400 grams,about 40 to 50% overlapping leaves inside the processing material of thehead, a short stem inside of the processing material of the head (i.e.,internal stem), leaves with about 4.8 to 5.3° Brix, leaves with about 25to 40 g/l total sugars, leaves with about 4.5 to 5% dry matter weight,5-10% processing damage per bag of processed plant material, and a rangeof about 35 to 42 days of shelf life of processed and packaged plantmaterial.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of a first iceberg lettuce plant with asecond iceberg lettuce plant, wherein the first iceberg lettuce plant isan upright heading iceberg lettuce plant including a space between abase of a head and a top of a ground, wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem), the second iceberg lettuce plant is an uprightheading iceberg lettuce plant including a space between a base of a headand a top of a ground, wherein said space includes an increased lengthof core outside of a processing material of a head (i.e., externalstem), and wherein the iceberg lettuce plant produced from the cross isan upright heading iceberg lettuce plant including a space between abase of a head and a top of a ground, wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem). In some embodiments, the first and second iceberglettuce plants are ‘E01E.70111’ Lot A lettuce plants, a sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957. In some embodiments, the first and secondiceberg lettuce plants are ‘E01E.70111’ Lot B lettuce plants, a sampleof ‘E01E.70111’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42962. In some embodiments, the first and secondiceberg lettuce plants are ‘E01E.70168’ Lot A lettuce plants, a sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958. In some embodiments, the first and secondiceberg lettuce plants are ‘E01E.70168’ Lot B lettuce plants, a sampleof ‘E01E.70168’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42963. In some embodiments, the first iceberg lettuceplant is an ‘E01E.70111’ Lot A lettuce plant, a sample of ‘E01E.70111’Lot A lettuce seed having been deposited under NCIMB Accession Number42957. In some embodiments, the second iceberg lettuce plant is an‘E01E.70168’ Lot A lettuce plant, a sample of ‘E01E.70168’ Lot A lettuceseed having been deposited under NCIMB Accession Number 42958.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot A plant with a second‘E01E.70111’ Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42957, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot B plant with a second‘E01E.70111’ Lot B plant, a sample of ‘E01E.70111’ Lot B lettuce seedhaving been deposited under NCIMB Accession Number 42962, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70168’ Lot A plant with a second‘E01E.70168’ Lot A plant, a sample of ‘E01E.70168’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42958, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70168’ Lot B plant with a second‘E01E.70168’ Lot B plant, a sample of ‘E01E.70168’ Lot B lettuce seedhaving been deposited under NCIMB Accession Number 42963, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot A plant with a second‘E01E.70168’ Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42957, and a sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958, wherein the iceberg lettuce plant produced fromthe cross includes a space between a base of a head and a top of aground, and wherein said space includes an increased length of coreoutside of a processing material of a head (i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceseed from the plants of any of the above embodiments. In anotherembodiment, the present disclosure is directed to iceberg lettuce plantsgrown from the seed of any of the above embodiments. In some embodimentsof any of the above embodiments, the iceberg lettuce plant includes aspace between a base of a head and a top of a ground, and said spaceincludes an increased length of core outside of a processing material ofa head (i.e., external stem). In some embodiments, the iceberg lettuceplant further includes one or more characteristics from the group of: aheight to diameter ratio of about 1.3 to about 1.5, leaf strengthbetween about 300 to 400 grams, about 40 to 50% overlapping leavesinside the processing material of the head, a short stem inside of theprocessing material of the head (i.e., internal stem), leaves with about4.8 to 5.3° Brix, leaves with about 25 to 40 g/l total sugars, leaveswith about 4.5 to 5% dry matter weight, 5-10% processing damage per bagof processed plant material, and a range of about 35 to 42 days of shelflife of processed and packaged plant material.

Lettuce plant parts include lettuce heads, lettuce leaves, parts oflettuce leaves, pollen, ovules, flowers, and the like. In anotherembodiment, the present disclosure is directed to a plant part of theplants of any of the above embodiments. In another embodiment, thepresent disclosure is further directed to heads, leaves, parts ofleaves, stems, roots, meristems, flowers, pollen, and ovules of theplants of any of the above embodiments. In another embodiment, thepresent disclosure is further directed to tissue culture of the plantsof any of the above embodiments, and to plants regenerated from thetissue culture, where the plant has all the morphological andphysiological characteristics of the plants of any of the aboveembodiments. In one embodiment, the plant regenerated from the tissueculture has all of the morphological and physiological characteristicsof an iceberg lettuce plant produced by growing seed designated as‘E01E.70111’ Lot A, representative sample of ‘E01E.70111’ Lot A lettuceseed having been deposited under NCIMB Accession Number 42957. Inanother embodiment, the plant regenerated from the tissue culture hasall of the morphological and physiological characteristics of an iceberglettuce plant produced by growing seed designated as ‘E01E.70111’ Lot B,representative sample of ‘E01E.70111’ Lot B lettuce seed having beendeposited under NCIMB Accession Number 42962. In one embodiment, theplant regenerated from the tissue culture has all of the morphologicaland physiological characteristics of an iceberg lettuce plant producedby growing seed designated as ‘E01E.70168’ Lot A, representative sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958. In another embodiment, the plant regeneratedfrom the tissue culture has all of the morphological and physiologicalcharacteristics of an iceberg lettuce plant produced by growing seeddesignated as ‘E01E.70168’ Lot B, representative sample of ‘E01E.70168’Lot B lettuce seed having been deposited under NCIMB Accession Number42963. In some embodiments, the present disclosure is directed to aplant part of any of the above embodiments of plants regenerated fromtissue culture. In some embodiments, the present disclosure is furtherdirected to heads, leaves, parts of leaves, stems, roots, meristems,flowers, pollen, and ovules of any of the above embodiments of plantsregenerated from tissue culture.

In still another embodiment, the present disclosure is further directedto packaging material containing plant parts of the plants of any of theabove embodiments. Such packaging material includes but is not limitedto boxes, plastic bags, etc. The plant parts of the plants of any of theabove embodiments may be combined with other plant parts of other plantvarieties.

In another embodiment, the present disclosure is directed to methods ofproducing an herbicide resistant lettuce plant by introducing a geneconferring herbicide resistance into a lettuce plant produced by growinglettuce seed of any of the above embodiments, where the gene is selectedfrom glyphosate, sulfonylurea, imidazolinone, dicamba, glufosinate,phenoxy proprionic acid, L-phosphinothricin, cyclohexone,cyclohexanedione, triazine, and benzonitrile. Certain embodiments arealso directed to herbicide resistant lettuce plants produced by suchmethods.

In another embodiment, the present disclosure is directed to methods ofproducing a pest or insect resistant lettuce plan by introducing a geneconferring pest or insect resistance into a lettuce plant produced bygrowing lettuce seed of any of the above embodiments, and to pest orinsect resistant lettuce plants produced by such methods. In certainembodiments, the gene conferring pest or insect resistance encodes aBacillus thuringiensis endotoxin.

In another embodiment, the present disclosure is directed to methods ofproducing a disease resistant lettuce plant by introducing a geneconferring disease resistance into a lettuce plant produced by growinglettuce seed of any of the above embodiments, and to disease resistantlettuce plants produced by such methods.

In another embodiment, the present disclosure is directed to methods ofproducing a lettuce plant with a value-added trait by introducing a geneconferring a value-added trait into a lettuce plant produced by growinglettuce seed of any of the above embodiments, where the gene encodes aprotein selected from a ferritin, a nitrate reductase, and a monellin.Certain embodiments are also directed to lettuce plants having avalue-added trait produced by such methods.

In another embodiment, the present disclosure provides for single geneconverted plants of any of the above embodiments. The single transferredgene may preferably be a dominant or recessive allele. Preferably, thesingle transferred gene will confer such traits as male sterility,herbicide resistance, insect or pest resistance, modified fatty acidmetabolism, modified carbohydrate metabolism, resistance for bacterial,fungal, or viral disease, male fertility, enhanced nutritional quality,and industrial usage. The single gene may be a naturally occurringlettuce gene.

In another embodiment, the present disclosure is directed to a method ofmaking iceberg lettuce seeds, by a) crossing the plant of any of theabove embodiments with another iceberg lettuce plant, and b) harvestingthe seed from the cross. In still another embodiment, the presentdisclosure is further directed to lettuce plants, lettuce parts from thelettuce plants, and seeds produced therefrom where the lettuce plant isisolated by the breeding method of the disclosure.

In yet another embodiment, the present disclosure is further directed toa method of making upright heading iceberg lettuce plants, by a)crossing a first upright heading iceberg lettuce plant including a spacebetween a base of a head and a top of a ground, wherein said spaceincludes an increased length of core outside of a processing material ofa head (i.e., external stem) with a second upright heading iceberglettuce plant including a space between a base of a head and a top of aground, wherein said space includes an increased length of core outsideof a processing material of a head (i.e., external stem); b) selectingoffspring lettuce plants including a space between a base of a head anda top of a ground, wherein said space includes an increased length ofcore outside of a processing material of a head (i.e., external stem);c) selfing or sibbing the offspring, and d) repeating steps b) and c)for multiple generations to produce inbred lines with the improved setof characteristics. In one embodiment, the characteristics used forselection in step b) further include one or more characteristics fromthe group of: a height to diameter ratio of about 1.3 to about 1.5, leafstrength between about 300 to 400 grams, about 40 to 50% overlappingleaves inside the processing material of the head, a short stem insideof the processing material of the head (i.e., internal stem), leaveswith about 4.8 to 5.3° Brix, leaves with about 25 to 40 g/l totalsugars, leaves with about 4.5 to 5% dry matter weight, 5-10% processingdamage per bag of processed plant material, and a range of about 35 to42 days of shelf life of processed and packaged plant material. In stillanother embodiment, the present disclosure is further directed tolettuce plants, lettuce parts from the lettuce plants, and seedsproduced therefrom where the lettuce plant is isolated by the breedingmethod of the disclosure.

In a further embodiment, the present disclosure relates to methods fordeveloping lettuce plants in a lettuce plant breeding program usingplant breeding techniques including recurrent selection, backcrossing,pedigree breeding, restriction fragment length polymorphism enhancedselection, genetic marker enhanced selection, and transformation. Seeds,lettuce plants, and parts thereof, produced by such breeding methods arealso part of the disclosure.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference bystudy of the following descriptions.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawings will be provided by the office upon request and paymentof the necessary fee.

FIGS. 1A & 1B show representative lettuce types. FIG. 1A shows anexemplary iceberg lettuce cross-section at the bottom left, and anexemplary upright heading iceberg at the center of the image. FIG. 1Bshows exemplary cosberg lettuce.

FIG. 2 shows visual quality scores of multiple lettuce varieties duringstorage.

FIG. 3 shows browning scores of multiple lettuce varieties.

FIG. 4 shows rotting scores of multiple lettuce varieties.

FIG. 5 shows intact leaves scores of multiple lettuce varietiesinitially and after 10 days of storage.

FIG. 6 shows the maturity index of iceberg lettuce.

FIG. 7A shows a cross-section of lettuce variety ‘E01E.70111’ Lot B.FIG. 7B shows a comparison of the bottom of ‘E01E.70111‘ Lot Aand’E01E.70111’ Lot B.

FIGS. 8A-8G show lettuce characteristics. FIG. 8A shows the degree ofoverlapping of the upper part of the leaves. FIG. 8B shows the number ofdivisions of the leaf. FIGS. 8C-8E show leaf shapes. FIG. 8F shows leafvenation. FIG. 8G shows head shape in longitudinal section.

FIGS. 9A-9J show exemplary whole heads and vertical cross-sections ofheads of lettuce varieties from Test 1. FIG. 9A shows an exemplary wholehead and vertical cross section of head of cosberg lettuce variety‘Cosmopolitan’ from Test 1. FIG. 9B shows an exemplary whole head andvertical cross section of head of cosberg lettuce variety ‘Crunchita’from Test 1. FIG. 9C shows an exemplary whole head and verticalcross-section of romaine lettuce variety ‘True Heart’ from Test 1.

FIG. 9D shows an exemplary whole head and vertical cross-section ofromaine lettuce variety ‘Solid King’ from Test 1. FIG. 9E shows anexemplary whole head and vertical cross-section of upright headingiceberg lettuce variety ‘E01E.70111’ Lot A from Test 1. FIG. 9F shows anexemplary whole head and vertical cross-section of upright headingiceberg lettuce variety ‘E01E.70111’ Lot B from Test 1. FIG. 9G shows anexemplary whole head and vertical cross-section of upright headingiceberg lettuce variety ‘E01E.70168’ Lot A from Test 1. FIG. 9H shows anexemplary whole head and vertical cross-section of upright headingiceberg lettuce variety ‘E01E.70168’ Lot B from Test 1. FIG. 9I shows anexemplary whole head and vertical cross-section of iceberg lettucevariety ‘Reliant’ from Test 1. FIG. 9J shows an exemplary whole head andvertical cross-section of iceberg lettuce variety ‘Steamboat’ from Test1.

FIGS. 10A-10J show exemplary whole heads and vertical cross-sections ofheads of lettuce varieties from Test 2. FIG. 10A shows an exemplarywhole head and vertical cross section of head of cosberg lettuce variety‘Cosmopolitan’ from Test 2. FIG. 10B shows an exemplary whole head andvertical cross section of head of cosberg lettuce variety ‘Crunchita’from Test 2. FIG. 10C shows an exemplary whole head and verticalcross-section of romaine lettuce variety ‘True Heart’ from Test 2. FIG.10D shows an exemplary whole head and vertical cross-section of romainelettuce variety ‘Solid King’ from Test 2. FIG. 10E shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70111’ Lot A from Test 2. FIG. 10F shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70111’ Lot B from Test 2. FIG. 10G shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70168’ Lot A from Test 2. FIG. 10H shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70168’ Lot B from Test 2. FIG. 10I shows an exemplarywhole head and vertical cross-section of iceberg lettuce variety‘Reliant’ from Test 2. FIG. 10J shows an exemplary whole head andvertical cross-section of iceberg lettuce variety ‘Steamboat’ from Test2.

FIGS. 11A-11J show exemplary whole heads and vertical cross-sections ofheads of lettuce varieties from Test 3. FIG. 11A shows an exemplarywhole head and vertical cross section of head of cosberg lettuce variety‘Cosmopolitan’ from Test 3. FIG. 11B shows an exemplary whole head andvertical cross section of head of cosberg lettuce variety ‘Crunchita’from Test 3. FIG. 11C shows an exemplary whole head and verticalcross-section of romaine lettuce variety ‘True Heart’ from Test 3. FIG.11D shows an exemplary whole head and vertical cross-section of romainelettuce variety ‘Solid King’ from Test 3. FIG. 11E shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70111’ Lot A from Test 3. FIG. 11F shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70111’ Lot B from Test 3. FIG. 11G shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70168’ Lot A from Test 3. FIG. 11H shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70168’ Lot B from Test 3. FIG. 11I shows an exemplarywhole head and vertical cross-section of iceberg lettuce variety‘Reliant’ from Test 3. FIG. 11J shows an exemplary whole head andvertical cross-section of iceberg lettuce variety ‘Steamboat’ from Test3.

FIGS. 12A-12J show exemplary whole heads and vertical cross-sections ofheads of lettuce varieties from Test 4. FIG. 12A shows an exemplarywhole head and vertical cross section of head of cosberg lettuce variety‘Cosmopolitan’ from Test 4. FIG. 12B shows an exemplary whole head andvertical cross section of head of cosberg lettuce variety ‘Crunchita’from Test 4. FIG. 12C shows an exemplary whole head and verticalcross-section of romaine lettuce variety ‘True Heart’ from Test 4. FIG.12D shows an exemplary whole head and vertical cross-section of romainelettuce variety ‘Solid King’ from Test 4. FIG. 12E shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70111’ Lot A from Test 4. FIG. 12F shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70111’ Lot B from Test 4. FIG. 12G shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70168’ Lot A from Test 4. FIG. 12H shows an exemplarywhole head and vertical cross-section of upright heading iceberg lettucevariety ‘E01E.70168’ Lot B from Test 4. FIG. 12I shows an exemplarywhole head and vertical cross-section of iceberg lettuce variety‘Reliant’ from Test 4. FIG. 12J shows an exemplary whole head andvertical cross-section of iceberg lettuce variety ‘Steamboat’ from Test4.

FIGS. 13A-13P show the stem length measurement procedure and theanalysis of stem length measurement data from Tests 1-4. FIG. 13A showsexemplary images of the external (left image) and internal (right image)stem length measurement procedures. External stem was measured from theharvest cut point at base of head to the first frame leaf of the head,and internal stem was measured from the first frame leaf of the head tothe end of the stem at the center of the head (shown in FIG. 29). FIGS.13B, 13E, 13H, 13K, and 13N show ANOVA analysis of the means of externaland internal stem length measurement data in cm from Test 1 (FIG. 13B),Test 2 (FIG. 13E), Test 3 (FIG. 13H), Test 4 (FIG. 13K), and all tests(i.e., Tests 1-4; FIG. 13N). For FIGS. 13B, 13E, 13H, 13K, and 13N, thebars depict the means, each lettuce variety is shown as adifferently-patterned bar (key at bottom of graph; from left to right,bars correspond to lettuce varieties ‘Cosmopolitan’, ‘Crunchita’, ‘TrueHeart’, ‘Solid King’, ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B,‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, ‘Reliant’, and ‘Steamboat’), andsignificantly different means between varieties are indicated bydifferent letters at the top of the bar. FIGS. 13C, 13F, 13I, 13L, and13O show box and whisker charts of external stem length data in cm fromTest 1 (FIG. 13C), Test 2 (FIG. 13F), Test 3 (FIG. 13I), Test 4 (FIG.13L), and all tests (i.e., Tests 1-4; FIG. 13O). FIGS. 13D, 13G, 13J,13M, and show box and whisker charts of internal stem length data in cmfrom Test 1 (FIG. 13D), Test 2 (FIG. 13G), Test 3 (FIG. 13J), Test 4(FIG. 13M), and all tests (i.e., Tests 1-4; FIG. 13P).

FIGS. 14A-14F show the lettuce head weight measurement procedure and theanalysis of lettuce head weight measurement data in grams (g) from Tests1-4. FIG. 14A shows an exemplary image of the lettuce head weightmeasurement procedure. FIGS. 14B-14F shows ANOVA analysis of the meansof lettuce head weight measurement in g from Test 1 (FIG. 14B), Test 2(FIG. 14C), Test 3 (FIG. 14D), Test 4 (FIG. 14E), and all tests (i.e.,Tests 1-4; FIG. 14F). For FIGS. 14B-14F, the bars depict the means, eachlettuce variety is shown as a differently-patterned bar (key at bottomof graph; from left to right, bars correspond to lettuce varieties‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘Solid King’, ‘E01E.70111’Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B,‘Reliant’, and ‘Steamboat’), and significantly different means betweenvarieties are indicated by different letters at the top of the bar.

FIGS. 15A-15F show the lettuce head height and lettuce head widthmeasurement procedures and the analysis of lettuce head height and widthmeasurement data in cm from Tests 1-4. FIG. 15A shows exemplary imagesof the lettuce head height (left image) and lettuce head width (rightimage) measurement procedures. FIGS. 15B-15F show ANOVA analysis of themeans of lettuce head height and width measurements in cm from Test 1(FIG. 15B), Test 2 (FIG. 15C), Test 3 (FIG. 15D), Test 4 (FIG. 15E), andall tests (i.e., Tests 1-4; FIG. 15F). For FIGS. 15B-15F, the barsdepict the means, each lettuce variety is shown as adifferently-patterned bar (key at bottom of graph; from left to right,bars correspond to lettuce varieties ‘Cosmopolitan’, ‘Crunchita’, ‘TrueHeart’, ‘Solid King’, ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B,‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, ‘Reliant’, and ‘Steamboat’), andsignificantly different means between varieties are indicated bydifferent letters at the top of the bar.

FIGS. 16A-16F show the evaluation of the percentage of overlappingleaves procedure and the analysis of the percentage of overlappingleaves data from Tests 1-4. FIG. 16A shows an exemplary image of alettuce head cut in half for use in the evaluation of the percentage ofoverlapping leaves procedure. FIGS. 16B-16F show ANOVA analysis of themeans of the percentage of overlapping leaves data from Test 1 (FIG.16B), Test 2 (FIG. 16C), Test 3 (FIG. 16D), Test 4 (FIG. 16E), and alltests (i.e., Tests 1-4; FIG. 16F). For FIGS. 16B-16F, the bars depictthe means, each lettuce variety is shown as a differently-patterned bar(key at bottom of graph; from left to right, bars correspond to lettucevarieties ‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘Solid King’,‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’Lot B, ‘Reliant’, and ‘Steamboat’), and significantly different meansbetween varieties are indicated by different letters at the top of thebar.

FIGS. 17A-17F show the leaf thickness measurement procedure and theanalysis of lettuce leaf thickness measurement data in millimeters (mm)from Tests 1-4. FIG. 17A shows exemplary images of the leaf thicknessmeasurement procedure. FIGS. 17B-17F show ANOVA analysis of the means ofleaf thickness measurement in mm from Test 1 (FIG. 17B), Test 2 (FIG.17C), Test 3 (FIG. 17D), Test 4 (FIG. 17E), and all tests (i.e., Tests1-4; FIG. 17F). For FIGS. 17B-17F, the bars depict the means, eachlettuce variety is shown as a differently-patterned bar (key at bottomof graph; from left to right, bars correspond to lettuce varieties‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘Solid King’, ‘E01E.70111’Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B,‘Reliant’, and ‘Steamboat’), and significantly different means betweenvarieties are indicated by different letters at the top of the bar.

FIGS. 18A-18P show the measurement of leaf color using thespectrophotometer procedure and the analysis of color lightness data,color hue angle data, and leaf gloss data from Tests 1-4. FIG. 18A showsexemplary images of the measurement of leaf color using thespectrophotometer procedure. FIGS. 18B-18F show ANOVA analysis of themeans of color lightness measurements from Test 1 (FIG. 18B), Test 2(FIG. 18C), Test 3 (FIG. 18D), Test 4 (FIG. 18E), and all tests (i.e.,Tests 1-4; FIG. 18F). FIGS. 18G-18K show ANOVA analysis of the means ofcolor hue angle measurements from Test 1 (FIG. 18G), Test 2 (FIG. 18H),Test 3 (FIG. 18I), Test 4 (FIG. 18J), and all tests (i.e., Tests 1-4;FIG. 18K). FIGS. 18L-18P show ANOVA analysis of the means of leaf glossmeasurements from Test 1 (FIG. 18L), Test 2 (FIG. 18M), Test 3 (FIG.18N), Test 4 (FIG. 18O), and all tests (i.e., Tests 1-4; FIG. 18P). ForFIGS. 18B-18P, the bars depict the means, each lettuce variety is shownas a differently-patterned bar (key at bottom of graph; from left toright, bars correspond to lettuce varieties ‘Cosmopolitan’, ‘Crunchita’,‘True Heart’, ‘Solid King’, ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B,‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, ‘Reliant’, and ‘Steamboat’), andsignificantly different means between varieties are indicated bydifferent letters at the top of the bar.

FIG. 19 shows an exemplary image of the measurement of leaf color usingthe RHS color chart procedure.

FIGS. 20A-20G show the measurement of leaf strength using thetexturometer procedure and the analysis of leaf strength measurementdata from Tests 1-4. FIGS. 20A-20B show exemplary images of themeasurement of leaf strength using the texturometer procedure. FIGS.20C-20G show ANOVA analysis of the means of measurement of leaf strengthusing the texturometer procedure from Test 1 (FIG. 20C), Test 2 (FIG.20D), Test 3 (FIG. 20E), Test 4 (FIG. 20F), and all tests (i.e., Tests1-4; FIG. 20G). For FIGS. 20C-20G, the bars depict the means, eachlettuce variety is shown as a differently-patterned bar (key at bottomof graph; from left to right, bars correspond to lettuce varieties‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘Solid King’, ‘E01E.70111’Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B,‘Reliant’, and ‘Steamboat’), and significantly different means betweenvarieties are indicated by different letters at the top of the bar.

FIG. 21 shows ANOVA analysis of the means of ° Brix measurements fromTests 1-4. The bars depict the means, each lettuce variety is shown as adifferently-patterned bar (key at bottom of graph; from left to right,bars correspond to lettuce varieties ‘Cosmopolitan’, ‘Crunchita’, ‘TrueHeart’, ‘Solid King’, ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B,‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, ‘Reliant’, and ‘Steamboat’), andsignificantly different means between varieties are indicated bydifferent letters at the top of the bar.

FIGS. 22A-22C show ANOVA analysis of the means of different sugarmeasurements across all tests (i.e., Tests 1-4). FIG. 22A shows ANOVAanalysis of the means of glucose, fructose, and sucrose measurements ing/l from all tests (i.e., Tests 1-4). FIG. 22B shows ANOVA analysis ofthe means of total sugar measurements in g/l from all tests (i.e., Tests1-4). FIG. 22C shows ANOVA analysis of the sweetness factor from alltests (i.e., Tests 1-4). For FIGS. 22A-22C, the bars depict the means,each lettuce variety is shown as a differently-patterned bar (key atbottom of graph; from left to right, bars correspond to lettucevarieties ‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘Solid King’,‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’Lot B, ‘Reliant’, and ‘Steamboat’), and significantly different meansbetween varieties are indicated by different letters at the top of thebar.

FIG. 23 shows ANOVA analysis of the means of percentage of dry matterweight measurements across all tests (i.e., Tests 1-4). The bars depictthe means, each lettuce variety is shown as a differently-patterned bar(key at bottom of graph; from left to right, bars correspond to lettucevarieties ‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘Solid King’,‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’Lot B, ‘Reliant’, and ‘Steamboat’), and significantly different meansbetween varieties are indicated by different letters at the top of thebar.

FIGS. 24A-24G show exemplary images of the plant material processingprocedure for shelf life evaluation. FIGS. 24A-24B show exemplary imagesof plant material cutting. FIG. 24C shows exemplary images of plantmaterial disinfection and drying. FIG. 24D shows an exemplary image ofplant material weighing and packaging. FIGS. 24E-24F show exemplaryimages of the process to achieve modified atmospheric conditions (MAP)and the sealed bags with MAP. FIG. 24G shows an exemplary image ofpackaged plant material in storage.

FIGS. 25A-25F show the analysis of the means of percentage of processingdamage data from Tests 1-4. FIGS. 25A, 25B, 25C, 25D, and 25F show boxand whisker charts of the percentage of processing damage data from Test1 (FIG. 25A), Test 2 (FIG. 25B), Test 3 (FIG. 25C), Test 4 (FIG. 25D),and all tests (i.e., Tests 1-4; FIG. 25F). FIG. 25E shows ANOVA analysisof the means of percentage of processing damage data from all tests(i.e., Tests 1-4). For FIG. 25E, the bars depict the means, each lettucevariety is shown as a differently-patterned bar (key at bottom of graph;from left to right, bars correspond to lettuce varieties ‘Cosmopolitan’,‘Crunchita’, ‘True Heart’, ‘Solid King’, ‘E01E.70111’ Lot A,‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, ‘Reliant’,and ‘Steamboat’), and significantly different means between varietiesare indicated by different letters at the top of the bar.

FIGS. 26A-26F show the analysis of the days of shelf life data fromTests 1-4. FIGS. 26A, 26B, 26C, 26D, and 26F show box and whisker chartsof the percentage of processing damage data from Test 1 (FIG. 26A), Test2 (FIG. 26B), Test 3 (FIG. 26C), Test 4 (FIG. 26D), and all tests (i.e.,Tests 1-4;

FIG. 26F). FIG. 26E shows ANOVA analysis of the means of percentage ofprocessing damage data from all tests (i.e., Tests 1-4). For FIG. 26E,the bars depict the means, each lettuce variety is shown as adifferently-patterned bar (key at bottom of graph; from left to right,bars correspond to lettuce varieties ‘Cosmopolitan’, ‘Crunchita’, ‘TrueHeart’, ‘Solid King’, ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B,‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, ‘Reliant’, and ‘Steamboat’), andsignificantly different means between varieties are indicated bydifferent letters at the top of the bar.

FIGS. 27A-27J show exemplary images of processed and packaged plantmaterial after 50 days of storage. FIG. 27A shows exemplary processedand packaged plant material outside and inside the packaging of cosberglettuce variety ‘Cosmopolitan’ after 50 days of storage. FIG. 27B showsexemplary processed and packaged plant material outside and inside thepackaging of cosberg lettuce variety ‘Crunchita’ after 50 days ofstorage. FIG. 27C shows exemplary processed and packaged plant materialoutside and inside the packaging of romaine lettuce variety ‘True Heart’after 50 days of storage. FIG. 27D shows exemplary processed andpackaged plant material outside and inside the packaging of romainelettuce variety ‘Solid King’ after 50 days of storage. FIG. 27E showsexemplary processed and packaged plant material outside and inside thepackaging of upright heading iceberg lettuce variety ‘E01E.70111’ Lot Aafter 50 days of storage. FIG. 27F shows exemplary processed andpackaged plant material outside and inside the packaging of uprightheading iceberg lettuce variety ‘E01E.70111’ Lot B after 50 days ofstorage. FIG. 27G shows exemplary processed and packaged plant materialoutside and inside the packaging of upright heading iceberg lettucevariety ‘E01E.70168’ Lot A after 50 days of storage. FIG. 27H showsexemplary processed and packaged plant material outside and inside thepackaging of upright heading iceberg lettuce variety ‘E01E.70168’ Lot Bafter 50 days of storage. FIG. 27I shows exemplary processed andpackaged plant material outside and inside the packaging of iceberglettuce variety ‘Reliant’ after 50 days of storage. FIG. 27J showsexemplary processed and packaged plant material outside and inside thepackaging of iceberg lettuce variety ‘Steamboat’ after 50 days ofstorage.

FIG. 28 shows a schematic of the machine cutting level on iceberg andupright heading iceberg lettuce varieties. Ground level is depicted by asolid line, machine cutting level is depicted by a dashed line, and thelocation of product loss (i.e., below the machine cutting level) foreach lettuce variety is labeled and indicated with a thick black arrow.

FIG. 29 shows a schematic of the external and internal core measurementson iceberg and upright heading iceberg varieties. Ground level isdepicted by a solid line, and cutting level is depicted by a dashedline. The processing material of the head is dark grey, and the stem(i.e., core) is light grey. The internal stem (i.e., core) is measuredabove the cutting level (labeled and indicated with a thick blackarrow), and the external stem (i.e., core) is measured below the cuttinglevel (labeled and indicated with a thick black arrow).

DETAILED DESCRIPTION

There are numerous steps in the development of novel, desirable lettucegermplasm. Plant breeding begins with the analysis of problems andweaknesses of current lettuce germplasms, the establishment of programgoals, and the definition of specific breeding objectives. The next stepis selection of germplasm that possess the traits to meet the programgoals. The goal is to combine in a single variety or hybrid an improvedcombination of desirable traits from the parental germplasm. Theseimportant traits may include increased head size and weight, higher seedyield, improved color, resistance to diseases and insects, tolerance todrought and heat, and better agronomic quality.

Choice of breeding or selection methods can depend on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of variety used commercially (e.g., F₁ hybrid variety, purelinevariety, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable variety. This approach hasbeen used extensively for breeding disease-resistant varieties. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program may include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, and can include gain fromselection per year based on comparisons to an appropriate standard, theoverall value of the advanced breeding lines, and the number ofsuccessful varieties produced per unit of input (e.g., per year, perdollar expended, etc.).

Promising advanced breeding lines may be thoroughly tested and comparedto appropriate standards in environments representative of thecommercial target area(s) for at least three years. The best lines canthen be candidates for new commercial varieties. Those still deficientin a few traits may be used as parents to produce new populations forfurther selection. These processes, which lead to the final step ofmarketing and distribution, may take from ten to twenty years from thetime the first cross or selection is made.

One goal of lettuce plant breeding is to develop new, unique andsuperior lettuce varieties. A breeder can initially select and cross twoor more parental lines, followed by repeated selfing and selection,producing many new genetic combinations. Moreover, a breeder cangenerate multiple different genetic combinations by crossing, selfing,and mutations. A plant breeder can then select which germplasms toadvance to the next generation. These germplasms may then be grown underdifferent geographical, climatic, and soil conditions, and furtherselections can be made during, and at the end of, the growing season.

The development of commercial lettuce varieties thus requires thedevelopment of parental lettuce varieties, the crossing of thesevarieties, and the evaluation of the crosses. Pedigree breeding andrecurrent selection breeding methods may be used to develop varietiesfrom breeding populations. Breeding programs can be used to combinedesirable traits from two or more varieties or various broad-basedsources into breeding pools from which new varieties are developed byselfing and selection of desired phenotypes. The new varieties arecrossed with other varieties and the hybrids from these crosses areevaluated to determine which have commercial potential.

Pedigree breeding is generally used for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents which possess favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁s or by intercrossing two F₁s (sib mating). Selection of the bestindividuals is usually begun in the F₂ population. Then, beginning inthe F₃, the best individuals in the best families are selected.Replicated testing of families, or hybrid combinations involvingindividuals of these families, often follows in the F₄ generation toimprove the effectiveness of selection for traits with low heritability.At an advanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new varieties.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding may be used to transfer genes for a simply inherited,highly heritable trait into a desirable homozygous cultivar or line thatis the recurrent parent. The source of the trait to be transferred iscalled the donor parent. The resulting plant is expected to have theattributes of the recurrent parent (e.g., cultivar) and the desirabletrait transferred from the donor parent. After the initial cross,individuals possessing the phenotype of the donor parent are selectedand repeatedly crossed (backcrossed) to the recurrent parent. Theresulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineswith each generation due to failure of some seeds to germinate or someplants to produce at least one seed. As a result, not all of the F₂plants originally sampled in the population will be represented by aprogeny when generation advance is completed.

In order to determine whether a lettuce plant is a plant of the presentdisclosure, and therefore whether said lettuce plant has the samegenetic determinant as plants of the present disclosure, the phenotypeof the lettuce plant can be compared with the phenotype of a known plantof the present disclosure. The phenotype can be assessed by, forexample, the ratio of height to diameter of the plant, the height of theplant, the crisp leaf texture, and/or the closed head.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. There are many laboratory-based techniques known in the artthat are available for the analysis, comparison and characterization ofplant genotype. Such techniques include, without limitation, IsozymeElectrophoresis, Restriction Fragment Length Polymorphisms (RFLPs),Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily PrimedPolymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs), AmplifiedFragment Length polymorphisms (AFLPs), Simple Sequence Repeats (SSRs,which are also referred to as Microsatellites), and Single NucleotidePolymorphisms (SNPs). By using these techniques, it is possible toassess the presence of the alleles, genes, and/or loci involved in theupright stature phenotype of the plants of the present disclosure. Forexample, performing linkage analysis between characteristics of apopulation (e.g. ‘E01E.70111’ Lot A) and genetic information of the samepopulation can provide markers linked to particular characteristics.This analysis can then be performed in offspring of the population tofurther confirm and improve the marker-characteristic linkage.

The genotype of a plant can also be used to determine the similaritybetween a plant of the present disclosure and a lettuce plant that maybe a plant of the present disclosure. For example, a marker from thegenotype of a plant of the present disclosure can be compared to amarker from the genotype of a lettuce plant that may be a plant of thepresent disclosure. The marker can be defined by a set of DNA-basedmarkers, such as AFLP, RFLP, RAPD, SCAR, CAPS, SSR, or SNP, which areclosely linked to the alleles, genes, or loci that are involved inproducing the upright stature phenotype of the present disclosure.Another method is to analyze the genotype of an F₂ population. In thismethod, first the plant of the present disclosure is crossed to theplant that may be a plant of the present disclosure. Then, the F₁ hybridoffspring of this cross is self-fertilized to produce the F₂ population.The phenotype and genotype of the F₂ population is then analyzed, forexample for the absence of segregation for the upright stature phenotypeand associated alleles, genes, or loci. In all of these geneticanalyses, phenotypic analyses can also be performed (e.g., by assessingthe ratio of height to diameter of the plant, the height of the plant,the crisp leaf texture, the closed head, etc.).

One procedure is to identify the equivalence between genetic informationresponsible for the upright stature phenotype of the plants of thepresent disclosure and the genetic information of a plant with anupright stature phenotype. First, the plant to be tested (with theupright stature phenotype, but unknown genotype) is used to makehomozygous offspring using methods known to the skilled person (e.g.,selfing) so that the genetic information responsible for the uprightstature phenotype will be homozygous. Then, the homozygous offspringplant is crossed with a tester plant that carries the geneticinformation responsible for the upright stature phenotype in homozygouscondition. If the plant to be tested has the upright stature phenotypeas a result of the same genetic information responsible for the uprightstature phenotype of plants of the present disclosure (i.e., the geneticinformation is equivalent), all progeny plants of this cross andsubsequent crosses will express the phenotype. If the upright staturephenotype of the plant to be tested is the result of a different part ofthe genome (i.e., the genetic information is not equivalent),segregation will occur in the population of offspring from the cross.The tester plant can be any plant that carries the genetic informationof the disclosure in homozygous condition, such as plants of whichrepresentative seed was deposited under accession number 42957, 42962,42958, or 42963, plants directly grown from the deposited seeds, orprogeny plants that have retained the phenotype.

The deposited seeds contain in their genome the genetic information thatencodes the upright stature phenotype. Therefore, the deposited seedsare a source for the genetic information that leads to the phenotype. Askilled person is capable of introducing the upright stature phenotypeinto any other plant using known techniques. Plants in the F₁ generation(i.e., the offspring from the cross between a first parent plant thatcontains the genetic information responsible for the upright staturephenotype with a second parent plant) may not be able to be identifiedas plants of the present disclosure (e.g., because of their heterozygouscondition). The phenotype is better assessed in an F₂ generation (e.g.,produced by selfing plants of the F₁ generation). One way of assessingthe phenotype would be to compare the stature of the F₂ progeny plantwith the second parent plant (e.g., by assessing the ratio of height todiameter of the plant, the height of the plant, etc.). If the stature ofthe F₂ progeny plant is more upright (e.g., has an increased ratio ofheight to diameter of the plant, an increased height of the plant, etc.)than that of the second parent plant, the progeny plant is a plant ofthe invention.

In another embodiment, the present disclosure provides plants that haveall of the morphological and physiological characteristics correspondingto the upright stature phenotype of plants of the present disclosure,representative seed of which having been deposited under NCIMB AccessionNumbers 42957, 42962, 42958, and 42963, which plants are grown fromseeds of a plant of the present disclosure, regenerated from partsthereof, or regenerated from a tissue culture thereof. Plants of thepresent disclosure should have the morphological and physiologicalcharacteristics that correspond with the upright stature phenotype butdo not necessarily have all the other characteristics of plants of thedeposited seeds. The upright stature phenotype is broadly transferableover multiple types and varieties.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select toward the genome of the recurrent parent and against themarkers of the donor parent. This procedure attempts to minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called genetic marker enhancedselection or marker-assisted selection. Molecular markers may also beused to identify and exclude certain sources of germplasm as parentalvarieties or ancestors of a plant by providing a means of trackinggenetic profiles through crosses.

Mutation breeding may also be used to introduce new traits into lettucevarieties. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation (such as X-rays, Gamma rays, neutrons,Beta radiation, or ultraviolet radiation), chemical mutagens (such asbase analogs like 5-bromo-uracil), antibiotics, alkylating agents (suchas sulfur mustards, nitrogen mustards, epoxides, ethyleneamines,sulfates, sulfonates, sulfones, or lactones), azide, hydroxylamine,nitrous acid, or acridines. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in Principles of Cultivar Development by Fehr, MacmillanPublishing Company (1993).

The production of double haploids can also be used for the developmentof homozygous varieties in a breeding program. Double haploids areproduced by the doubling of a set of chromosomes from a heterozygousplant to produce a completely homozygous individual. For example, seeWan, et al., Theor. Appl. Genet., 77:889-892 (1989).

Additional non-limiting examples of breeding methods that may be usedinclude, without limitation, those found in Principles of PlantBreeding, John Wiley and Son, pp. 115-161 (1960); Allard (1960);Simmonds (1979); Sneep, et al. (1979); Fehr (1987); and “Carrots andRelated Vegetable Umbelliferae,” Rubatzky, V. E., et al. (1999).

Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. The allele is any of one or more alternative forms of a gene,all of which relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Big Vein virus. Big vein is a disease of lettuce caused by LettuceMirafiori Big Vein Virus which is transmitted by the fungus Olpidiumvirulentus, with vein clearing and leaf shrinkage resulting in plants ofpoor quality and reduced marketable value.

Bolting. The premature development of a flowering stalk, and subsequentseed, before a plant produces a food crop. Bolting is typically causedby late planting.

Bremia lactucae. An oomycete that causes downy mildew in lettuce incooler growing regions.

Core length. Length of the internal lettuce stem measured from the baseof the cut and trimmed head to the tip of the stem.

Corky root. A disease caused by the bacterium Sphingomonassuberifaciens, which causes the entire taproot to become brown, severelycracked, and non-functional.

Cotyledon. One of the first leaves of the embryo of a seed plant;typically one or more in monocotyledons, two in dicotyledons, and two ormore in gymnosperms.

Essentially all the physiological and morphological characteristics. Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having the physiological and morphologicalcharacteristics of the recurrent parent, except for the characteristicsderived from the converted gene.

External stem length: External stem (i.e., core) was measured from theharvest cut point at the base of the head the first frame leaf of thehead. The location of the external stem (i.e., core) is shown in FIG.29.

First water date. The date the seed first receives adequate moisture togerminate. This can and often does equal the planting date.

Frame leaves. The leaves that form the base of the plant. Generally notconsidered a part of the processing material (e.g., head).

Gene. As used herein, “gene” refers to a segment of nucleic acid. A genecan be introduced into a genome of a species, whether from a differentspecies or from the same species, using transformation or variousbreeding methods.

Head diameter. Diameter of the cut and trimmed head, sliced vertically,and measured at the widest point perpendicular to the stem.

Head height. Height of the cut and trimmed head, sliced vertically, andmeasured from the base of the cut stem to the cap leaf.

Head weight. Weight of saleable lettuce head, cut and trimmed to marketspecifications.

Internal stem length: Internal stem (i.e., core) was measured from thefirst frame leaf of the head to the end of the stem at the center of thehead. The location of the internal stem (i.e., core) is shown in FIG.29.

Lettuce Mosaic virus. A disease that can cause a stunted, deformed, ormottled pattern in young lettuce and yellow, twisted, and deformedleaves in older lettuce.

Maturity date. Maturity refers to the stage when the plants are of fullsize or optimum weight, in marketable form or shape to be of commercialor economic value.

Nasonovia ribisnigri. A lettuce aphid that colonizes the innermostleaves of the lettuce plant, contaminating areas that cannot be treatedeasily with insecticides.

Quantitative Trait Loci. Quantitative Trait Loci (QTL) refers to geneticloci that control to some degree, numerically representable traits thatare usually continuously distributed.

Ratio of head height/diameter. Head height divided by the head diameteris an indication of the head shape; <1 is flattened, 1=round, and >1 ispointed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Rhizoctonia solani. A soil-borne fungus that causes bottom rot inlettuce.

RHS. RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd., RHS Garden;Wisley, Woking; Surrey GU236QB, UK.

Single gene converted. Single gene converted or conversion plant refersto plants which are developed by a plant breeding technique calledbackcrossing or via genetic engineering where essentially all of thedesired morphological and physiological characteristics of a line arerecovered in addition to the single gene transferred into the line viathe backcrossing technique or via genetic engineering.

Tipburn. Means a browning of the edges or tips of lettuce leaves that isa physiological response to a lack of calcium.

Wet date. The wet date corresponds to the first planting date oflettuce.

Upright Heading Iceberg Lettuce Plants and Seeds

Certain aspects of the present disclosure relate to upright headingiceberg plants, and to seeds that produce the upright heading iceberglettuce plants described herein.

Plants

Certain aspects of the present disclosure relate to an upright headingiceberg lettuce plant with a height to diameter ratio greater than orequal to about 1.0, about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head. In someembodiments, the height of the standard iceberg lettuce plant is about 5inches to 6 inches. In some embodiments, the height of the uprightheading iceberg lettuce plant is about 7.5 inches to about 18 inches. Insome embodiments, a height of a head of an upright heading iceberglettuce plant is about 1.5 to about 3 times a height of a head of astandard iceberg lettuce plant. In some embodiments, the height of thehead of the standard iceberg lettuce plant is about 5 inches to 6inches. In some embodiments, the height of the head of the uprightheading iceberg lettuce plant is about 7.5 inches to about 18 inches. Insome embodiments, the head does not include frame leaves. In someembodiments, the height of the upright heading lettuce plant isdependent on the season in which the upright heading lettuce plant isgrown (e.g., winter, spring, summer, fall). In some embodiments, thediameter of the upright heading lettuce plant is dependent on the seasonin which the upright heading lettuce plant is grown. In someembodiments, the height to diameter ratio is dependent on the season inwhich the upright heading lettuce plant is grown. In some embodiments,the upright heading lettuce plant may not grow as tall due tosub-optimal growing conditions or a poorly grown field. In someembodiments, the crisp leaf texture is the result of high water content.In some embodiments, the crisp leaf texture is the result of thickleaves. In some embodiments, the crisp leaf texture is due to acombination of factors (e.g., high water content and thick leaves). Insome embodiments, the crisp leaf texture is a characteristic thatenables specific processing types (e.g., chopping). In some embodiments,the crisp leaf texture is a subjective assessment of texture and/or apleasant sensation when chewing.

In some embodiments, the iceberg lettuce plant further includes one ormore characteristics from the group of: a short core, a low level ofinternal tipburn, a low level of bottom rot, an increased number ofleaves, a fast fill rate of leaves in a head, and a space between a baseof a head and a top of a ground. In some embodiments, thecharacteristics above only apply to the processing material (e.g. head).In some embodiments, the characteristics above do not apply to frameleaves. In some embodiments, the short core is short inside the head. Insome embodiments, the short core is short outside the head. In someembodiments, core length is the same as in iceberg, romaine, or cosbergtypes. In some embodiments, core length is shorter than in iceberg,romaine, or cosberg types. In some embodiments, a low level of tipburnmeans so no visible symptoms. In some embodiments, tipburn is the sameas in iceberg, romaine, or cosberg types. In some embodiments, tipburnis less than in iceberg, romaine, or cosberg types. In some embodiments,the level of tipburn is dependent on external conditions (e.g.,environment, growing process, etc.). For example, in some embodiments,the upright heading iceberg lettuce plant is about 2 to about 3 timestaller than a standard iceberg lettuce plant, has a similar core lengthto a standard iceberg lettuce plant, and has tipburn tolerance. In someembodiments, a low level of bottom rot means no visible symptoms. Insome embodiments, the level of bottom rot is the same as in icebergtypes. In some embodiments, the level of bottom rot is less than iniceberg types. In some embodiments, the level of bottom rot is dependenton external conditions (e.g., environment, growing process, etc.). Insome embodiments, less bottom rot is due to less contact of the headwith the ground. In some embodiments, a low level of bottom rot is dueto less contact of the head with the ground than in iceberg types. Insome embodiments, the problems associated with bottom rot are avoidedbecause the harvest machine cuts higher on the upright heading icebergplant than the standard iceberg lettuce plant. In some embodiments, anincreased number of leaves means an increased number of leaves in thehead. In some embodiments, the iceberg lettuce plant produces anincreased number of frame leaves. In some embodiments, an increasednumber of frame leaves results in the processing material being higheroff of the ground. In some embodiments, the fill rate is slower than thefill rate of iceberg lettuce. In some embodiments, the fill rate issimilar to the fill rate of iceberg lettuce. In some embodiments, a fastfill rate of leaves in a head is comparable to the fill rate of aniceberg lettuce. In some embodiments, a fast fill rate of leaves isfaster than the fill rate of an iceberg lettuce. In some embodiments, afast fill rate of leaves in a head results in tightly packed leaves atharvest time. In some embodiments, tightly packed leaves at harvest timeresult in blanched inner leaves that are paler in color than the outerleaves. In some embodiments, a fast fill rate of leaves in a head meansthat the core remains short and tipburn is reduced. In some embodiments,the space between the base of the head and the top of the ground isachieved by an increased length of core outside of the processingmaterial of a head. In some embodiments, the space between the base ofthe head and the top of the ground is sufficient for machine harvesting.In some embodiments, the base of the head is a similar distance to theground as iceberg lettuce, the head is taller, and the bottom of thehead is cut off during harvesting (e.g., machine harvesting). In someembodiments, a small proportion of processing material is cut off (i.e.,lost) during harvest (e.g., machine harvest). In some embodiments,upright heading iceberg lettuce varieties allow processing of the sameamount of product per acre with a machine as can be processed forstandard iceberg lettuce varieties using hand labor. In someembodiments, the use of a machine allows significant cost savings incomparison to hand labor. In some embodiments, the processor will beable to harvest less upright heading iceberg lettuce product per acrethan standard iceberg lettuce product, and will still be able to make aprofit. In some embodiments, the iceberg lettuce plant further has thecharacteristic of good shelf life, where good shelf life means similarshelf life qualities as a standard iceberg lettuce. In some embodiments,shelf life encompasses odor, rotting, and browning or pinking.

An exemplary upright heading iceberg lettuce plant is illustrated inFIG. 1A. A longitudinal cross-section of the upright heading iceberglettuce plant (FIG. 1A center) is shown next to a longitudinalcross-section of an iceberg lettuce plant (FIG. 1A bottom left). Withoutwishing to be bound by theory, it is thought that the heading and leafdensity of the upright heading iceberg lettuce plant is similar to thatof the iceberg lettuce plant. Without wishing to be bound by theory, itis further thought that the upright heading iceberg lettuce plant isboth taller and narrower than the iceberg lettuce plant, which means agreater amount of processing material is further away from the ground,and thus the upright heading iceberg lettuce plant is machineharvestable. Exemplary cosberg plants are depicted in FIG. 1B. Withoutwishing to be bound by theory, it is thought that cosberg plants aregenerally shorter in stature than upright heading iceberg lettuceplants, not generally suitable for mechanical harvest, and not able tobe used to replace product from standard iceberg plants. Without wishingto be bound by theory, it is further thought that cosberg plants do nottaste like standard iceberg plants, and also do not have the same crisptexture as standard iceberg plants.

In some embodiments, the iceberg lettuce plant further includes one ormore characteristics from the group of: moderate leaf glossiness,moderate tendency to bolt, inner leaves paler than outer leaves,blanched inner leaves, green to greyish green outer leaves, outer leaveswith a color ranging from about RHS146A to about RHS146B, inner leaveswith a color ranging from RHS 145C to RHS 245D, leaves with an obovateshape, leaves with an elliptic shape, leaves with a broad obtrullateshape, leaves with a triangular shape, head with a narrow elliptic shapein longitudinal section, head with an elliptic shape in longitudinalsection, head with a broad elliptic shape in longitudinal section, headwith an oval shape in longitudinal section, head with an oblong shape inlongitudinal section, medium degree of overlapping of upper part ofleaves, strong degree of overlapping of upper part of leaves, absent orvery few leaf divisions, leaf margin hardly to rather strongly incised,thick leaves, absent to medium undulation of leaf margin,semi-flabellate leaf venation, flabellate leaf venation, clear midrib,no clear midrib, heading, early heading, and medium heading. In otherembodiments, the iceberg lettuce plant further includes one or morecharacteristics from the group of characteristics defining the uprightheading iceberg lettuce plant type. Exemplary upright heading iceberglettuce characteristics are illustrated in FIGS. 8A-8G. FIG. 8Aillustrates the appearance of different degrees of overlapping of upperparts of leaves, including medium to strong degrees of overlapping. FIG.8B illustrates the appearance of leaf divisions, including absent orvery few leaf divisions. FIGS. 8C-8E illustrate different leaf shapes,including obovate, elliptic, and broad obtrullate. FIG. 8F illustratesleaf venation patterns, including flabellate venation. FIG. 8Gillustrates the shape of the head in longitudinal section, includingnarrow elliptic and broad elliptic. For example, in some embodiments, anupright heading lettuce plant of the current disclosure may have amedium degree of overlapping of upper parts of leaves, absent or veryfew leaf divisions, obovate leaves, flabellate leaf venation, and anarrow elliptic shape of the head in longitudinal section. For example,in some embodiments, an upright heading lettuce plant of the currentdisclosure may have a strong degree of overlapping of upper parts ofleaves, absent or very few leaf divisions, obovate leaves, flabellateleaf venation, and a broad elliptic shape of the head in longitudinalsection.

In some embodiments, the height to diameter ratio of an upright headingiceberg plant of the present disclosure is greater than or equal toabout 1.0, greater than or equal to about 1.1, greater than or equal toabout 1.2, greater than or equal to about 1.3, greater than or equal toabout 1.4, greater than or equal to about 1.5, greater than or equal toabout 1.6, greater than or equal to about 1.7, greater than or equal toabout 1.8, greater than or equal to about 1.9, greater than or equal toabout 2.0, greater than or equal to about 2.1, greater than or equal toabout 2.2, greater than or equal to about 2.3, greater than or equal toabout 2.4, greater than or equal to about 2.5, greater than or equal toabout 2.6, greater than or equal to about 2.7, greater than or equal toabout 2.8, greater than or equal to about 2.9, greater than or equal toabout 3.0, greater than or equal to about 3.1, greater than or equal toabout 3.2, greater than or equal to about 3.3, greater than or equal toabout 3.4, or greater than or equal to about 3.5. In some embodiments,the height to diameter ratio of an upright heading iceberg plant of thepresent disclosure is less than or equal to about 3.5, less than orequal to about 3.4, less than or equal to about 3.3, less than or equalto about 3.2, less than or equal to about 3.1, less than or equal toabout 3.0, less than or equal to about 2.9, less than or equal to about2.8, less than or equal to about 2.7, less than or equal to about 2.6,less than or equal to about 2.5, less than or equal to about 2.4, lessthan or equal to about 2.3, less than or equal to about 2.2, less thanor equal to about 2.1, less than or equal to about 2.0, less than orequal to about 1.9, less than or equal to about 1.8, less than or equalto about 1.7, less than or equal to about 1.6, less than or equal toabout 1.5, less than or equal to about 1.4, less than or equal to about1.3, less than or equal to about 1.2, less than or equal to about 1.1,or less than or equal to about 1.0. In some embodiments, the height todiameter ratio of an upright heading iceberg plant of the presentdisclosure is greater than or equal to about any of the followingratios: 1.0, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6,2.7, 2.8, 2.9, 3.0, or 3.5. In some embodiments, the height to diameterratio of an upright heading iceberg plant of the present disclosure isless than or equal to about any of the following ratios: 3.5, 3.0, 2.9,2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, or1.0. In some embodiments, the height to diameter ratio of an uprightheading iceberg plant of the present disclosure can be any of a range ofratios having a lower limit of 1.0, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1,2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, or 3.5 and an independentlyselected upper limit of 3.5, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3,2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, or 1.0. For example, in someembodiments, the height to diameter ratio of an upright heading iceberglettuce plant of the present disclosure is between about 1.5 and 3.0.

In some embodiments, the height of an upright heading iceberg plant ofthe present disclosure is greater than or equal to about 7.5 in, greaterthan or equal to about 8 in, greater than or equal to about 8.5 in,greater than or equal to about 9 in, greater than or equal to about 9.5in, greater than or equal to about 10 in, greater than or equal to about10.5 in, greater than or equal to about 11 in, greater than or equal toabout 11.5 in, greater than or equal to about 12 in, greater than orequal to about 12.5 in, greater than or equal to about 13 in, greaterthan or equal to about 13.5 in, greater than or equal to about 14 in,greater than or equal to about 14.5 in, greater than or equal to about15 in, greater than or equal to about 15.5 in, greater than or equal toabout 16 in, greater than or equal to about 16.5 in, greater than orequal to about 17 in, greater than or equal to about 17.5 in, greaterthan or equal to about 18 in, greater than or equal to about 19 in,greater than or equal to about 20 in, greater than or equal to about 21in, greater than or equal to about 22 in, greater than or equal to about23 in, greater than or equal to about 24 in, or greater than or equal toabout 25 in. In some embodiments, the height of an upright headingiceberg plant of the present disclosure is less than or equal to about25 in, less than or equal to about 24 in, less than or equal to about 23in, less than or equal to about 22 in, less than or equal to about 21in, less than or equal to about 20 in, less than or equal to about 19in, less than or equal to about 18 in, less than or equal to about 17.5in, less than or equal to about 17 in, less than or equal to about 16.5in, less than or equal to about 16 in, less than or equal to about 15.5in, less than or equal to about 15 in, less than or equal to about 14.5in, less than or equal to about 14 in, less than or equal to about 13.5in, less than or equal to about 13 in, less than or equal to about 12.5in, less than or equal to about 12 in, less than or equal to about 11.5in, less than or equal to about 11 in, less than or equal to about 10.5in, less than or equal to about 10 in, less than or equal to about 9.5in, less than or equal to about 9 in, less than or equal to about 8.5in, less than or equal to about 8 in, or less than or equal to about 7.5in. In some embodiments, the height of an upright heading iceberg plantof the present disclosure is greater than or equal to about any of thefollowing heights: 7.5 in, 8 in, 9 in, 10 in, 11 in, 12 in, 13 in, 14in, 15 in, 16 in, 17 in, 18 in, 20 in, or 25 in. In some embodiments,the height of an upright heading iceberg plant of the present disclosureis less than or equal to about any of the following heights: 25 in, 20in, 18 in, 17 in, 16 in, 15 in, 14 in, 13 in, 12 in, 11 in, 10 in, 9 in,8 in, 7.5 in. In some embodiments, the height of an upright headingiceberg plant of the present disclosure can be any of a range of heightshaving a lower limit of 7.5 in, 8 in, 9 in, 10 in, 11 in, 12 in, 13 in,14 in, 15 in, 16 in, 17 in, 18 in, 20 in, or 25 in and an independentlyselected upper limit of 5. For example, in some embodiments, the heightof an upright heading iceberg lettuce plant of the present disclosure isbetween about 7.5 in and about 18 in.

In some embodiments, an iceberg lettuce plant is produced from a crossof a first iceberg lettuce plant with a second iceberg lettuce plant,wherein the first iceberg lettuce plant produces an upright headingiceberg lettuce plant with a height to diameter ratio greater than orequal to about 1.0, about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head, thesecond iceberg lettuce plant produces an upright heading iceberg lettuceplant with a height to diameter ratio greater than or equal to about1.0, about 1.5 to about 3 times a height of a standard iceberg lettuceplant, a crisp leaf texture, and a closed head, and wherein the iceberglettuce plant produced from the cross has an upright heading iceberglettuce plant with a height to diameter ratio greater than or equal toabout 1.0, a height of about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head. In someembodiments, the iceberg lettuce plant produced from the cross has aheight to diameter ratio greater than or equal to about 1.5, a heightgreater than or equal to about 25.0 cm, and a closed head. In someembodiments, the first and second iceberg lettuce plants are‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, or‘E01E.70168’ Lot B lettuce plants, samples of each having been depositedunder NCIMB Accession Numbers 42957, 42962, 42958, or 42963respectively. In some embodiments, only one of the iceberg lettuceplants (i.e, the first or the second iceberg lettuce plant) is‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, or‘E01E.70168’ Lot B lettuce plants, samples of each having been depositedunder NCIMB Accession Numbers 42957, 42962, 42958, or 42963respectively. In one embodiment, the first lettuce plant and the secondplant are from different lines (e.g., ‘E01E.70111‘ Lot A and’E01E.70111’Lot B). In another embodiment, the first lettuce plant and the secondplant are from the same line (e.g., ‘E01E.70111’ Lot A). In someembodiments, the plants produced from these crosses have an uprightheading iceberg lettuce plant with a height to diameter ratio greaterthan or equal to about 1.0, a height of about 1.5 to about 3 times aheight of a standard iceberg lettuce plant, a crisp leaf texture, and aclosed head.

In another embodiment, the present disclosure relates to one or moreplant parts from any of the upright heading iceberg lettuce plantsdescribed herein. In some embodiments, the plant part is a head, a leaf,a seed, a cell, or any portion thereof. In some embodiments, the plantpart is a head. In some embodiments, the head is upright heading. Insome embodiments, the head has the characteristics of a height todiameter ratio greater than or equal to about 1.0, a height of about 1.5to about 3 times a height of a standard iceberg lettuce plant, a crispleaf texture, and a closed head. In some embodiments, the plant part isa stem, a root, a meristem, or a flower.

In some embodiments, the present disclosure relates to one or morepollen grains or one or more ovules from any of the upright headingiceberg lettuce plants described herein. In some embodiments, thepresent disclosure relates to a pollen grain from any of the uprightheading iceberg lettuce plants described herein. In some embodiments,the present disclosure relates to an ovule from any of the uprightheading iceberg lettuce plants described herein.

In some embodiments, the present disclosure relates to tissue cultureproduced from protoplasts or cells from any of the upright headingiceberg lettuce plants described herein. In some embodiments, theprotoplasts and/or cells are produced from one or more plant parts fromany of the upright heading iceberg lettuce plants described herein. Insome embodiments, the plant part is one or more of leaf, anther, pistil,core, stem, root, root tip, flower, seed, cotyledon, hypocotyl, embryo,and meristematic cells. In some embodiments, the plants regenerated fromthe tissue culture have all the morphological and physiologicalcharacteristics of the plants of any of the upright heading iceberglettuce plants described herein. In one embodiment, the plantregenerated from the tissue culture has all of the morphological andphysiological characteristics of an iceberg lettuce plant produced bygrowing seed designated as ‘E01E.70111’ Lot A, representative sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957. In another embodiment, the plant regeneratedfrom the tissue culture has all of the morphological and physiologicalcharacteristics of an iceberg lettuce plant produced by growing seeddesignated as ‘E01E.70111’ Lot B, representative sample of ‘E01E.70111’Lot B lettuce seed having been deposited under NCIMB Accession Number42962. In one embodiment, the plant regenerated from the tissue culturehas all of the morphological and physiological characteristics of aniceberg lettuce plant produced by growing seed designated as‘E01E.70168’ Lot A, representative sample of ‘E01E.70168’ Lot A lettuceseed having been deposited under NCIMB Accession Number 42958. Inanother embodiment, the plant regenerated from the tissue culture hasall of the morphological and physiological characteristics of an iceberglettuce plant produced by growing seed designated as ‘E01E.70168’ Lot B,representative sample of ‘E01E.70168’ Lot B lettuce seed having beendeposited under NCIMB Accession Number 42963.

In certain aspects, the present disclosure relates to an upright headingiceberg lettuce plant including a space between a base of a head and atop of a ground (e.g., ground level). In some embodiments, said spaceincludes an increased length of core outside of a processing material ofa head (i.e., external stem). In some embodiments, the increased lengthof core outside of the processing material of the head (i.e., externalstem) is about 3.5 cm or more, about 4 cm or more, about 4.5 cm or more,or about 5 cm or more. In some embodiments, the iceberg lettuce plantfurther includes one or more, two or more, three or more, four or more,five or more, six or more, seven or more, eight or more, or nine or morecharacteristics selected from the group of: a height to diameter ratioof about 1.3 to about 1.5, leaf strength between about 300 to 400 grams,about 40 to 50% overlapping leaves inside the processing material of thehead, a short stem inside of the processing material of the head (i.e.,internal stem), leaves with about 4.8 to 5.3° Brix, leaves with about 25to 40 g/l total sugars, leaves with about 4.5 to 5% dry matter weight,5-10% processing damage per bag of processed plant material, and a rangeof about 35 to 42 days of shelf life of processed and packaged plantmaterial.

In some embodiments, the increased length of core outside of theprocessing material of the head (i.e., external stem) of an uprightheading iceberg plant of the present disclosure is greater than or equalto about 3.0 cm, greater than or equal to about 3.1 cm, greater than orequal to about 3.2 cm, greater than or equal to about 3.3 cm, greaterthan or equal to about 3.4 cm, greater than or equal to about 3.5 cm,greater than or equal to about 3.6 cm, greater than or equal to about3.7 cm, greater than or equal to about 3.8 cm, greater than or equal toabout 3.9 cm, greater than or equal to about 4.0 cm, greater than orequal to about 4.1 cm, greater than or equal to about 4.2 cm, greaterthan or equal to about 4.3 cm, greater than or equal to about 4.4 cm,greater than or equal to about 4.5 cm, greater than or equal to about4.6 cm, greater than or equal to about 4.7 cm, greater than or equal toabout 4.8 cm, greater than or equal to about 4.9 cm, greater than orequal to about 5.0 cm, greater than or equal to about 5.1 cm, greaterthan or equal to about 5.2 cm, greater than or equal to about 5.3 cm,greater than or equal to about 5.4 cm, or greater than or equal to about5.5 cm. In some embodiments, the increased length of core outside of theprocessing material of the head (i.e., external stem) of an uprightheading iceberg plant of the present disclosure is less than or equal toabout 5.5 cm, less than or equal to about 5.4 cm, less than or equal toabout 5.3 cm, less than or equal to about 5.2 cm, less than or equal toabout 5.1 cm, less than or equal to about 5.0 cm, less than or equal toabout 4.9 cm, less than or equal to about 4.8 cm, less than or equal toabout 4.7 cm, less than or equal to about 4.6 cm, less than or equal toabout 4.5 cm, less than or equal to about 4.4 cm, less than or equal toabout 4.3 cm, less than or equal to about 4.2 cm, less than or equal toabout 4.1 cm, less than or equal to about 4.0 cm, less than or equal toabout 3.9 cm, less than or equal to about 3.8 cm, less than or equal toabout 3.7 cm, less than or equal to about 3.6 cm, less than or equal toabout 3.5 cm, less than or equal to about 3.4 cm, less than or equal toabout 3.3 cm, less than or equal to about 3.2 cm, less than or equal toabout 3.1 cm, or less than or equal to about 3.0 cm. In someembodiments, the increased length of core outside of the processingmaterial of the head (i.e., external stem) of an upright heading icebergplant of the present disclosure is greater than or equal to thefollowing values in cm: 3.5, 3.7, 4.0, 4.2, 4.5, 4.7, or 5.0. In someembodiments, the increased length of core outside of the processingmaterial of the head (i.e., external stem) of an upright heading icebergplant of the present disclosure is less than or equal to about any ofthe following values in cm: 5.0, 4.7, 4.5, 4.2, 4.0, 3.7, or 3.5. Insome embodiments, the increased length of core outside of the processingmaterial of the head (i.e., external stem) of an upright heading icebergplant of the present disclosure can be any of a range of values in cmhaving a lower limit of 3.5, 3.7, 4.0, 4.2, 4.5, 4.7, or 5.0 and anindependently selected upper limit of 5.0, 4.7, 4.5, 4.2, 4.0, 3.7, or3.5. For example, in some embodiments, the increased length of coreoutside of the processing material of the head (i.e., external stem) ofan upright heading iceberg lettuce plant of the present disclosure isbetween about 3.5 and 5.0 cm.

In some embodiments, the height to diameter ratio of an upright headingiceberg plant of the present disclosure is greater than or equal toabout 1.20, greater than or equal to about 1.21, greater than or equalto about 1.22, greater than or equal to about 1.23, greater than orequal to about 1.24, greater than or equal to about 1.25, greater thanor equal to about 1.26, greater than or equal to about 1.27, greaterthan or equal to about 1.28, greater than or equal to about 1.29,greater than or equal to about 1.3, greater than or equal to about 1.31,greater than or equal to about 1.32, greater than or equal to about1.33, greater than or equal to about 1.34, greater than or equal toabout 1.35, greater than or equal to about 1.36, greater than or equalto about 1.37, greater than or equal to about 1.38, greater than orequal to about 1.39, greater than or equal to about 1.40, greater thanor equal to about 1.41, greater than or equal to about 1.42, greaterthan or equal to about 1.43, greater than or equal to about 1.44,greater than or equal to about 1.45, greater than or equal to about1.46, greater than or equal to about 1.47, greater than or equal toabout 1.48, greater than or equal to about 1.49, greater than or equalto about 1.50, greater than or equal to about 1.51, greater than orequal to about 1.52, greater than or equal to about 1.53, greater thanor equal to about 1.54, greater than or equal to about 1.55, greaterthan or equal to about 1.56, greater than or equal to about 1.57,greater than or equal to about 1.58, greater than or equal to about1.59, or greater than or equal to about 1.60. In some embodiments, theheight to diameter ratio of an upright heading iceberg plant of thepresent disclosure is less than or equal to about 1.60, less than orequal to about 1.59, less than or equal to about 1.58, less than orequal to about 1.57, less than or equal to about 1.56, less than orequal to about 1.55, less than or equal to about 1.54, less than orequal to about 1.53, less than or equal to about 1.52, less than orequal to about 1.51, less than or equal to about 1.50, less than orequal to about 1.49, less than or equal to about 1.48, less than orequal to about 1.47, less than or equal to about 1.46, less than orequal to about 1.45, less than or equal to about 1.44, less than orequal to about 1.43, less than or equal to about 1.42, less than orequal to about 1.41, less than or equal to about 1.40, less than orequal to about 1.39, less than or equal to about 1.38, less than orequal to about 1.37, less than or equal to about 1.36, less than orequal to about 1.35, less than or equal to about 1.34, less than orequal to about 1.33, less than or equal to about 1.32, less than orequal to about 1.31, less than or equal to about 1.30, less than orequal to about 1.29, less than or equal to about 1.28, less than orequal to about 1.27, less than or equal to about 1.26, less than orequal to about 1.25, less than or equal to about 1.24, less than orequal to about 1.23, less than or equal to about 1.22, less than orequal to about 1.21, or less than or equal to about 1.20. In someembodiments, the height to diameter ratio of an upright heading icebergplant of the present disclosure is greater than or equal to about any ofthe following ratios: 1.30, 1.31, 1.32, 1.33, 1.34, 1.35, 1.36, 1.37,1.38, 1.39, 1.40, 1.41, 1.42, 1.43, 1.44, 1.45, 1.46, 1.47, 1.48, 1.49,or 1.50. In some embodiments, the height to diameter ratio of an uprightheading iceberg plant of the present disclosure is less than or equal toabout any of the following ratios: 1.50, 1.49, 1.48, 1.47, 1.46, 1.45,1.44, 1.43, 1.42, 1.41, 1.40, 1.39, 1.38, 1.37, 1.36, 1.35, 1.34, 1.33,1.32, 1.31, or 1.30. In some embodiments, the height to diameter ratioof an upright heading iceberg plant of the present disclosure can be anyof a range of ratios having a lower limit of 1.30, 1.31, 1.32, 1.33,1.34, 1.35, 1.36, 1.37, 1.38, 1.39, 1.40, 1.41, 1.42, 1.43, 1.44, 1.45,1.46, 1.47, 1.48, 1.49, or 1.50 and an independently selected upperlimit of 1.50, 1.49, 1.48, 1.47, 1.46, 1.45, 1.44, 1.43, 1.42, 1.41,1.40, 1.39, 1.38, 1.37, 1.36, 1.35, 1.34, 1.33, 1.32, 1.31, or 1.30. Forexample, in some embodiments, the height to diameter ratio of an uprightheading iceberg lettuce plant of the present disclosure is between about1.3 and 1.5.

In some embodiments, the leaf strength of an upright heading icebergplant of the present disclosure is greater than or equal to about 250grams, greater than or equal to about 260 grams, greater than or equalto about 270 grams, greater than or equal to about 280 grams, greaterthan or equal to about 290 grams, greater than or equal to about 300grams, greater than or equal to about 305 grams, greater than or equalto about 310 grams, greater than or equal to about 315 grams, greaterthan or equal to about 320 grams, greater than or equal to about 325grams, greater than or equal to about 330 grams, greater than or equalto about 335 grams, greater than or equal to about 340 grams, greaterthan or equal to about 345 grams, greater than or equal to about 350grams, greater than or equal to about 355 grams, greater than or equalto about 360 grams, greater than or equal to about 365 grams, greaterthan or equal to about 370 grams, greater than or equal to about 375grams, greater than or equal to about 380 grams, greater than or equalto about 385 grams, greater than or equal to about 390 grams, greaterthan or equal to about 395 grams, greater than or equal to about 400grams, greater than or equal to about 410 grams, greater than or equalto about 420 grams, greater than or equal to about 430 grams, greaterthan or equal to about 440 grams, greater than or equal to about 450grams, greater than or equal to about 460 grams, greater than or equalto about 470 grams, greater than or equal to about 480 grams, greaterthan or equal to about 490 grams, or greater than or equal to about 500grams. In some embodiments, the leaf strength of an upright headingiceberg plant of the present disclosure is less than or equal to about500 grams, less than or equal to about 490 grams, less than or equal toabout 480 grams, less than or equal to about 470 grams, less than orequal to about 460 grams, less than or equal to about 450 grams, lessthan or equal to about 440 grams, less than or equal to about 430 grams,less than or equal to about 420 grams, less than or equal to about 410grams, less than or equal to about 400 grams, less than or equal toabout 395 grams, less than or equal to about 390 grams, less than orequal to about 385 grams, less than or equal to about 380 grams, lessthan or equal to about 375 grams, less than or equal to about 370 grams,less than or equal to about 365 grams, less than or equal to about 360grams, less than or equal to about 355 grams, less than or equal toabout 350 grams, less than or equal to about 345 grams, less than orequal to about 340 grams, less than or equal to about 335 grams, lessthan or equal to about 330 grams, less than or equal to about 325 grams,less than or equal to about 320 grams, less than or equal to about 315grams, less than or equal to about 310 grams, less than or equal toabout 305 grams, less than or equal to about 300 grams, less than orequal to about 290 grams, less than or equal to about 280 grams, lessthan or equal to about 270 grams, less than or equal to about 260 grams,less than or equal to about 250 grams, less than or equal to about 240grams, less than or equal to about 230 grams, less than or equal toabout 220 grams, less than or equal to about 210 grams, less than orequal to about 210 grams, or less than or equal to about 200 grams. Insome embodiments, the leaf strength of an upright heading iceberg plantof the present disclosure is greater than or equal to about any of thefollowing values in grams: 300, 310, 320, 330, 340, 350, 360, 370, 380,390, or 400. In some embodiments, the leaf strength of an uprightheading iceberg plant of the present disclosure is greater than or equalto about any of the following values in grams: 400, 390, 380, 370, 360,350, 340, 330, 320, 310, or 300. In some embodiments, the leaf strengthof an upright heading iceberg plant of the present disclosure can be anyof a range of values in grams having a lower limit of 300, 310, 320,330, 340, 350, 360, 370, 380, 390, or 400 and an independently selectedupper limit of 400, 390, 380, 370, 360, 350, 340, 330, 320, 310, or 300.For example, in some embodiments, the leaf strength of an uprightheading iceberg lettuce plant of the present disclosure is between about300 and about 400 grams.

In some embodiments, the percent of overlapping leaves of an uprightheading iceberg plant of the present disclosure is greater than or equalto about 35%, greater than or equal to about 36%, greater than or equalto about 37%, greater than or equal to about 38%, greater than or equalto about 39%, greater than or equal to about 40%, greater than or equalto about 41%, greater than or equal to about 42%, greater than or equalto about 43%, greater than or equal to about 44%, greater than or equalto about 45%, greater than or equal to about 46%, greater than or equalto about 47%, greater than or equal to about 48%, greater than or equalto about 49%, greater than or equal to about 50%, greater than or equalto about 51%, greater than or equal to about 52%, greater than or equalto about 53%, greater than or equal to about 54%, or greater than orequal to about 55%. In some embodiments, the percent of overlappingleaves of an upright heading iceberg plant of the present disclosure isless than or equal to about 55%, less than or equal to about 54%, lessthan or equal to about 53%, less than or equal to about 52%, less thanor equal to about 51%, less than or equal to about 50%, less than orequal to about 49%, less than or equal to about 48%, less than or equalto about 47%, less than or equal to about 46%, less than or equal toabout 45%, less than or equal to about 44%, less than or equal to about43%, less than or equal to about 42%, less than or equal to about 41%,less than or equal to about 40%, less than or equal to about 39%, lessthan or equal to about 38%, less than or equal to about 37%, less thanor equal to about 36%, or less than or equal to about 35%. In someembodiments, the percent of overlapping leaves of an upright headingiceberg plant of the present disclosure is greater than or equal toabout any of the following percentages: 40%, 41%, 42%, 43%, 44%, 45%,46%, 47%, 48%, 49%, or 50%. In some embodiments, the percent ofoverlapping leaves of an upright heading iceberg plant of the presentdisclosure is less than or equal to about any of the followingpercentages: 50%, 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, or 40%.In some embodiments, the percent of overlapping leaves of an uprightheading iceberg plant of the present disclosure can be any of a range ofpercentages having a lower limit of 40%, 41%, 42%, 43%, 44%, 45%, 46%,47%, 48%, 49%, or 50% and an independently selected upper limit of 50%,49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, or 40%. For example, insome embodiments, the percent of overlapping leaves of an uprightheading iceberg lettuce plant of the present disclosure is between about40% and 50%.

In some embodiments, the ° brix of leaves of an upright heading icebergplant of the present disclosure is greater than or equal to about 4.5°brix, greater than or equal to about 4.6° brix, greater than or equal toabout 4.7° brix, greater than or equal to about 4.8° brix, greater thanor equal to about 4.85° brix, greater than or equal to about 4.9° brix,greater than or equal to about 4.95° brix, greater than or equal toabout 5.0° brix, greater than or equal to about 5.05° brix, greater thanor equal to about 5.1° brix, greater than or equal to about 5.15° brix,greater than or equal to about 5.2° brix, greater than or equal to about5.25° brix, greater than or equal to about 5.3° brix, greater than orequal to about 5.4° brix, greater than or equal to about 5.5° brix,greater than or equal to about 5.6° brix, or greater than or equal toabout 5.7° brix. In some embodiments, the ° brix of leaves of an uprightheading iceberg plant of the present disclosure is less than or equal toabout 5.7° brix, less than or equal to about 5.5° brix, less than orequal to about 5.4° brix, less than or equal to about 5.3° brix, lessthan or equal to about 5.25° brix, less than or equal to about 5.2°brix, less than or equal to about 5.15° brix, less than or equal toabout 5.1° brix, less than or equal to about 5.05° brix, less than orequal to about 5.0° brix, less than or equal to about 4.95° brix, lessthan or equal to about 4.9° brix, less than or equal to about 4.85°brix, less than or equal to about 4.8° brix, less than or equal to about4.7° brix, less than or equal to about 4.6° brix, or less than or equalto about 4.6° brix. In some embodiments, the ° brix of leaves of anupright heading iceberg plant of the present disclosure is greater thanor equal to about any of the following values in ° brix: 4.8, 4.85, 4.9,4.95, 5.0, 5.05, 5.1, 5.15, 5.2, 5.25, or 5.3. In some embodiments, the° brix of an upright heading iceberg plant of the present disclosure isgreater than or equal to about any of the following values in ° brix:5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0, 4.95, 4.9, 4.85, or 4.8. In someembodiments, the ° brix of leaves of an upright heading iceberg plant ofthe present disclosure can be any of a range of values in ° brix havinga lower limit of 4.8, 4.85, 4.9, 4.95, 5.0, 5.05, 5.1, 5.15, 5.2, 5.25,or 5.3 and an independently selected upper limit of 5.3, 5.25, 5.2,5.15, 5.1, 5.05, 5.0, 4.95, 4.9, 4.85, or 4.8. For example, in someembodiments, the ° brix of leaves of an upright heading iceberg lettuceplant of the present disclosure is between about 4.8° brix and about5.3° brix.

In some embodiments, the total sugars of leaves of an upright headingiceberg plant of the present disclosure are greater than or equal toabout 20 g/l, greater than or equal to about 21 g/l, greater than orequal to about 22 g/l, greater than or equal to about 23 g/l, greaterthan or equal to about 24 g/l, greater than or equal to about 25 g/l,greater than or equal to about 26 g/l, greater than or equal to about 27g/l, greater than or equal to about 28 g/l, greater than or equal toabout 29 g/l, greater than or equal to about 30 g/l, greater than orequal to about 31 g/l, greater than or equal to about 32 g/l, greaterthan or equal to about 33 g/l, greater than or equal to about 34 g/l,greater than or equal to about 35 g/l, greater than or equal to about 36g/l, greater than or equal to about 37 g/l, greater than or equal toabout 38 g/l, greater than or equal to about 39 g/l, greater than orequal to about 40 g/l, greater than or equal to about 41 g/l, greaterthan or equal to about 42 g/l, greater than or equal to about 43 g/l,greater than or equal to about 44 g/l, or greater than or equal to about45 g/l. In some embodiments, the total sugars of leaves of an uprightheading iceberg plant of the present disclosure are less than or equalto about 45 g/l, less than or equal to about 44 g/l, less than or equalto about 43 g/l, less than or equal to about 42 g/l, less than or equalto about 41 g/l, less than or equal to about 40 g/l, less than or equalto about 39 g/l, less than or equal to about 38 g/l, less than or equalto about 37 g/l, less than or equal to about 36 g/l, less than or equalto about 35 g/l, less than or equal to about 34 g/l, less than or equalto about 33 g/l, less than or equal to about 32 g/l, less than or equalto about 31 g/l, less than or equal to about 30 g/l, less than or equalto about 29 g/l, less than or equal to about 28 g/l, less than or equalto about 27 g/l, less than or equal to about 26 g/l, less than or equalto about 25 g/l, less than or equal to about 24 g/l, less than or equalto about 23 g/l, less than or equal to about 22 g/l, less than or equalto about 21 g/l, or less than or equal to about 20 g/l. In someembodiments, the total sugars of leaves of an upright heading icebergplant of the present disclosure are greater than or equal to about anyof the following values in g/l: 25, 27, 30, 32, 35, 37, or 40. In someembodiments, the total sugars of leaves of an upright heading icebergplant of the present disclosure are greater than or equal to about anyof the following values in g/l: 40, 37, 35, 32, 30, 27, or 25. In someembodiments, the total sugars of leaves of an upright heading icebergplant of the present disclosure can be any of a range of values in g/lhaving a lower limit of 25, 27, 30, 32, 35, 37, or 40 and anindependently selected upper limit of 40, 37, 35, 32, 30, 27, or 25. Forexample, in some embodiments, the total sugars of leaves of an uprightheading iceberg lettuce plant of the present disclosure are betweenabout 25 g/l and about 40 g/l.

In some embodiments, the percent of dry matter weight of an uprightheading iceberg plant of the present disclosure is greater than or equalto about 4.0%, greater than or equal to about 4.1%, greater than orequal to about 4.2%, greater than or equal to about 4.3%, greater thanor equal to about 4.4%, greater than or equal to about 4.5%, greaterthan or equal to about 4.55%, greater than or equal to about 4.6%,greater than or equal to about 4.65%, greater than or equal to about4.7%, greater than or equal to about 4.75%, greater than or equal toabout 4.8%, greater than or equal to about 4.85%, greater than or equalto about 4.9%, greater than or equal to about 4.95%, greater than orequal to about 5.0%, greater than or equal to about 5.1%, greater thanor equal to about 5.2%, greater than or equal to about 5.3%, greaterthan or equal to about 5.4%, or greater than or equal to about 5.5%. Insome embodiments, the percent of dry matter weight of an upright headingiceberg plant of the present disclosure is less than or equal to about5.5%, less than or equal to about 5.4%, less than or equal to about5.3%, less than or equal to about 5.2%, less than or equal to about5.1%, less than or equal to about 5.0%, less than or equal to about4.95%, less than or equal to about 4.9%, less than or equal to about4.85%, less than or equal to about 4.8%, less than or equal to about4.75%, less than or equal to about 4.7%, less than or equal to about4.65%, less than or equal to about 4.6%, less than or equal to about4.55%, less than or equal to about 4.5%, less than or equal to about4.4%, less than or equal to about 4.3%, less than or equal to about4.2%, less than or equal to about 4.1%, or less than or equal to about4.0%. In some embodiments, the percent of dry matter weight of anupright heading iceberg plant of the present disclosure is greater thanor equal to about any of the following percentages: 4.5%, 4.55%, 4.6%,4.65%, 4.7%, 4.75%, 4.8%, 4.85%, 4.9%, 4.95%, or 5.0%. In someembodiments, the percent of dry matter weight of an upright headingiceberg plant of the present disclosure is less than or equal to aboutany of the following percentages: 5.0%, 4.95%, 4.9%, 4.85%, 4.8%, 4.75%,4.7%, 4.65%, 4.6%, 4.55%, or 4.5%. In some embodiments, the percent ofdry matter weight of an upright heading iceberg plant of the presentdisclosure can be any of a range of percentages having a lower limit of4.5%, 4.55%, 4.6%, 4.65%, 4.7%, 4.75%, 4.8%, 4.85%, 4.9%, 4.95%, or 5.0%and an independently selected upper limit of 5.0%, 4.95%, 4.9%, 4.85%,4.8%, 4.75%, 4.7%, 4.65%, 4.6%, 4.55%, or 4.5%. For example, in someembodiments, the percent of dry matter weight of an upright headingiceberg lettuce plant of the present disclosure is between about 4.5%and 5%.

In some embodiments, the processing damage per bag of processed plantmaterial of an upright heading iceberg plant of the present disclosureis greater than or equal to about 4.0%, greater than or equal to about0%, greater than or equal to about 1%, greater than or equal to about2%, greater than or equal to about 3%, greater than or equal to about4%, greater than or equal to about 5%, greater than or equal to about6%, greater than or equal to about 7%, greater than or equal to about8%, greater than or equal to about 9%, greater than or equal to about10%, greater than or equal to about 11%, or greater than or equal toabout 12%. In some embodiments, the processing damage per bag ofprocessed plant material of an upright heading iceberg plant of thepresent disclosure is less than or equal to about 12%, less than orequal to about 11%, less than or equal to about 10%, less than or equalto about 9%, less than or equal to about 8%, less than or equal to about7%, less than or equal to about 6%, less than or equal to about 5%, lessthan or equal to about 4%, less than or equal to about 3%, less than orequal to about 2%, less than or equal to about 1%, or less than or equalto about 0%. In some embodiments, the processing damage per bag ofprocessed plant material of an upright heading iceberg plant of thepresent disclosure is greater than or equal to about any of thefollowing percentages: 5%, 6%, 7%, 8%, 9%, or 10%. In some embodiments,the processing damage per bag of processed plant material of an uprightheading iceberg plant of the present disclosure is less than or equal toabout any of the following percentages: 10%, 9%, 8%, 7%, 6%, or 5%. Insome embodiments, the processing damage per bag of processed plantmaterial of an upright heading iceberg plant of the present disclosurecan be any of a range of percentages having a lower limit of %, 6%, 7%,8%, 9%, or 10% and an independently selected upper limit of 10%, 9%, 8%,7%, 6%, or 5%. For example, in some embodiments, the processing damageper bag of processed plant material of an upright heading iceberglettuce plant of the present disclosure is between about 5% and 10%.

In some embodiments, the days of shelf life of processed and packagedplant material of an upright heading iceberg plant of the presentdisclosure are greater than or equal to about greater than or equal toabout 30 days, greater than or equal to about 31 days, greater than orequal to about 32 days, greater than or equal to about 33 days, greaterthan or equal to about 34 days, greater than or equal to about 35 days,greater than or equal to about 36 days, greater than or equal to about37 days, greater than or equal to about 38 days, greater than or equalto about 39 days, greater than or equal to about 40 days, greater thanor equal to about 41 days, greater than or equal to about 42 days,greater than or equal to about 43 days, greater than or equal to about44 days, greater than or equal to about 45 days, greater than or equalto about 46 days, greater than or equal to about 47 days, greater thanor equal to about 48 days, greater than or equal to about 49 days, orgreater than or equal to about 50 days. In some embodiments, the days ofshelf life of processed and packaged plant material of an uprightheading iceberg plant of the present disclosure are less than or equalto about 50 days, less than or equal to about 49 days, less than orequal to about 48 days, less than or equal to about 47 days, less thanor equal to about 46 days, less than or equal to about 45 days, lessthan or equal to about 44 days, less than or equal to about 43 days,less than or equal to about 42 days, less than or equal to about 41days, less than or equal to about 40 days, less than or equal to about39 days, less than or equal to about 38 days, less than or equal toabout 37 days, less than or equal to about 36 days, less than or equalto about 35 days, less than or equal to about 34 days, less than orequal to about 33 days, less than or equal to about 32 days, less thanor equal to about 31 days, or less than or equal to about 30 days. Insome embodiments, the days of shelf life of processed and packaged plantmaterial of an upright heading iceberg plant of the present disclosureare greater than or equal to about any of the following values in days:35, 36, 37, 38, 39, 40, 41, or 42. In some embodiments, the days ofshelf life of processed and packaged plant material of an uprightheading iceberg plant of the present disclosure are greater than orequal to about any of the following values in days: 42, 41, 40, 39, 38,37, 36, or 35. In some embodiments, the days of shelf life of processedand packaged plant material of an upright heading iceberg plant of thepresent disclosure can be any of a range of values in days having alower limit of 35, 36, 37, 38, 39, 40, 41, or 42 and an independentlyselected upper limit of 42, 41, 40, 39, 38, 37, 36, or 35. For example,in some embodiments, the days of shelf life of processed and packagedplant material of an upright heading iceberg lettuce plant of thepresent disclosure are between about 35 days and about 42 days.

Certain aspects of the present disclosure relate to an iceberg lettuceseed, wherein the seed produces an iceberg lettuce plant with an uprightstature phenotype. In some embodiments, the upright stature phenotypeincludes a space between a base of a head and a top of a ground (e.g.,ground level). In some embodiments, said space includes an increasedlength of core outside of a processing material of a head (i.e.,external stem). In some embodiments, the increased length of coreoutside of the processing material of the head (i.e., external stem) isabout 3.5 cm or more, about 4 cm or more, about 4.5 cm or more, or about5 cm or more. In some embodiments, the upright stature phenotype furtherincludes one or more characteristics selected from the group of: aheight to diameter ratio of about 1.3 to about 1.5, leaf strengthbetween about 300 to 400 grams, about 40 to 50% overlapping leavesinside the processing material of the head, a short stem inside of theprocessing material of the head (i.e., internal stem), leaves with about4.8 to 5.3° Brix, leaves with about 25 to 40 g/l total sugars, leaveswith about 4.5 to 5% dry matter weight, 5-10% processing damage per bagof processed plant material, and a range of about 35 to 42 days of shelflife of processed and packaged plant material.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of a first iceberg lettuce plant with asecond iceberg lettuce plant, wherein the first iceberg lettuce plant isan upright heading iceberg lettuce plant including a space between abase of a head and a top of a ground, wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem), the second iceberg lettuce plant is an uprightheading iceberg lettuce plant including a space between a base of a headand a top of a ground, wherein said space includes an increased lengthof core outside of a processing material of a head (i.e., externalstem), and wherein the iceberg lettuce plant produced from the cross isan upright heading iceberg lettuce plant including a space between abase of a head and a top of a ground, wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem). In some embodiments, the first and second iceberglettuce plants are ‘E01E.70111’ Lot A lettuce plants, a sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957. In some embodiments, the first and secondiceberg lettuce plants are ‘E01E.70111’ Lot B lettuce plants, a sampleof ‘E01E.70111’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42962. In some embodiments, the first and secondiceberg lettuce plants are ‘E01E.70168’ Lot A lettuce plants, a sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958. In some embodiments, the first and secondiceberg lettuce plants are ‘E01E.70168’ Lot B lettuce plants, a sampleof ‘E01E.70168’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42963. In some embodiments, the first iceberg lettuceplant is an ‘E01E.70111’ Lot A lettuce plant, a sample of ‘E01E.70111’Lot A lettuce seed having been deposited under NCIMB Accession Number42957. In some embodiments, the second iceberg lettuce plant is an‘E01E.70168’ Lot A lettuce plant, a sample of ‘E01E.70168’ Lot A lettuceseed having been deposited under NCIMB Accession Number 42958.

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot A plant with a second‘E01E.70111’ Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42957, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot B plant with a second‘E01E.70111’ Lot B plant, a sample of ‘E01E.70111’ Lot B lettuce seedhaving been deposited under NCIMB Accession Number 42962, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70168’ Lot A plant with a second‘E01E.70168’ Lot A plant, a sample of ‘E01E.70168’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42958, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70168’ Lot B plant with a second‘E01E.70168’ Lot B plant, a sample of ‘E01E.70168’ Lot B lettuce seedhaving been deposited under NCIMB Accession Number 42963, wherein theiceberg lettuce plant produced from the cross includes a space between abase of a head and a top of a ground, and wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem).

Certain aspects of the present disclosure relate to an iceberg lettuceplant produced from a cross of an ‘E01E.70111’ Lot A plant with a second‘E01E.70168’ Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42957, and a sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958, wherein the iceberg lettuce plant produced fromthe cross includes a space between a base of a head and a top of aground, and wherein said space includes an increased length of coreoutside of a processing material of a head (i.e., external stem).

In some embodiments that may be combined with any of the aboveembodiments, the external stem length allows for a harvest by machine.In some embodiments, the harvest by machine does not remove asubstantial portion of processing material of a head. In someembodiments, the harvest by machine is more efficient (i.e., removesless processing material of the head) than for iceberg varieties‘Reliant’ and ‘Steamboat’, cosberg varieties ‘Cosmopolitan’ and‘Crunchita’, and romaine varieties ‘True Heart’ and ‘Solid King’.

Lettuce plant parts include lettuce heads, lettuce leaves, parts oflettuce leaves, pollen, ovules, flowers, and the like. In anotherembodiment, the present disclosure is directed to a plant part of theplants of any of the above embodiments. In another embodiment, thepresent disclosure is further directed to heads, leaves, parts ofleaves, stems, roots, meristems, flowers, pollen, and ovules of theplants of any of the above embodiments. In another embodiment, thepresent disclosure is further directed to tissue culture of the plantsof any of the above embodiments, and to plants regenerated from thetissue culture, where the plant has all the morphological andphysiological characteristics of the plants of any of the aboveembodiments. In one embodiment, the plant regenerated from the tissueculture has all of the morphological and physiological characteristicsof an iceberg lettuce plant produced by growing seed designated as‘E01E.70111’ Lot A, representative sample of ‘E01E.70111’ Lot A lettuceseed having been deposited under NCIMB Accession Number 42957. Inanother embodiment, the plant regenerated from the tissue culture hasall of the morphological and physiological characteristics of an iceberglettuce plant produced by growing seed designated as ‘E01E.70111’ Lot B,representative sample of ‘E01E.70111’ Lot B lettuce seed having beendeposited under NCIMB Accession Number 42962. In one embodiment, theplant regenerated from the tissue culture has all of the morphologicaland physiological characteristics of an iceberg lettuce plant producedby growing seed designated as ‘E01E.70168’ Lot A, representative sampleof ‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958. In another embodiment, the plant regeneratedfrom the tissue culture has all of the morphological and physiologicalcharacteristics of an iceberg lettuce plant produced by growing seeddesignated as ‘E01E.70168’ Lot B, representative sample of ‘E01E.70168’Lot B lettuce seed having been deposited under NCIMB Accession Number42963. In some embodiments, the present disclosure is directed to aplant part of any of the above embodiments of plants regenerated fromtissue culture. In some embodiments, the present disclosure is furtherdirected to heads, leaves, parts of leaves, stems, roots, meristems,flowers, pollen, and ovules of any of the above embodiments of plantsregenerated from tissue culture.

Seeds

Certain aspects of the present disclosure relate to one or more seedsthat produce upright heading iceberg lettuce plants described herein.Certain aspects of the present disclosure relate to one or more seeds,wherein the seed produces an iceberg lettuce plant with an uprightstature phenotype. In some embodiments, the upright stature phenotypeincludes a height to diameter ratio greater than or equal to about 1.0,a height of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head. In someembodiments, the iceberg lettuce plant further includes one or morecharacteristics from the group of: a short core, a low level of internaltipburn, a low level of bottom rot, an increased number of leaves, afast fill rate of leaves in a head, and a space between a base of a headand a top of a ground.

Certain aspects of the present disclosure relate to an iceberg lettuceseed containing an upright stature allele at locus A, wherein the seedproduces upright heading iceberg lettuce plants described herein.Certain aspects of the present disclosure relate to an iceberg lettuceseed comprising an upright stature allele at locus A, wherein the seedproduces an iceberg lettuce with an upright stature phenotype. In someembodiments, the upright stature phenotype includes a height to diameterratio greater than or equal to about 1.0, a height of about 1.5 to about3 times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head. In another embodiment, the iceberg lettucefurther includes one or more characteristics from the group of: a shortcore, a low level of internal tipburn, a low level of bottom rot, anincreased number of leaves, a fast fill rate of leaves in a head, and aspace between a base of a head and a top of a ground.

The upright heading iceberg lettuce plants of the present disclosure maybe produced from seed containing an upright stature allele (e.g. uprightstature allele at locus A). In some embodiments, the allele is dominant.In some embodiments, the allele is recessive. In some embodiments, theallele is semi-dominant. In some embodiments, more than one allelecontributes to the upright stature phenotype (e.g., a multi-gene trait,a polygenic trait, etc.). In some embodiments, the expression of one ormore alleles is upregulated to produce the upright stature phenotype. Insome embodiments, the expression of one or more alleles is downregulatedto produce the upright stature phenotype. In some embodiments, theupright stature phenotype is the result of gene expression andenvironmental conditions (e.g., heat). In some embodiments, the uprightstature phenotype is the result of gene expression and cultivationconditions (e.g., close planting). In some embodiments, specificcharacteristics of upright stature can be linked to specific alleles(e.g., increased height to height allele at locus B). In someembodiments, linked markers can be used to identify these alleles.

Certain aspects of the present disclosure relate to one or more seedsthat produce upright heading iceberg lettuce plants described herein.Certain aspects of the present disclosure relate to an iceberg lettuceseed comprising an upright stature allele at locus A, wherein the seedproduces upright heading iceberg lettuce plants described herein.Certain aspects of the present disclosure relate to an iceberg lettuceseed comprising an upright stature allele at locus A, wherein the seedproduces an iceberg lettuce with an upright stature phenotype. In oneembodiment, the upright stature phenotype includes a height to diameterratio greater than or equal to about 1.0, a height of about 1.5 to about3 times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head. In another embodiment, the iceberg lettucefurther includes one or more characteristics from the group of: a shortcore, a low level of internal tipburn, a low level of bottom rot, anincreased number of leaves, a fast fill rate of leaves in a head, and aspace between a base of a head and a top of a ground.

The upright heading iceberg lettuce plants of the present disclosure maybe produced from seed containing an upright stature allele (e.g. uprightstature allele at locus A). In some embodiments, the allele is dominant.In some embodiments, the allele is recessive. In some embodiments, theallele is semi-dominant. In some embodiments, more than one allelecontributes to the upright stature phenotype (e.g., a multi-gene trait,a polygenic trait, etc.). In some embodiments, the expression of one ormore alleles is upregulated to produce the upright stature phenotype. Insome embodiments, the expression of one or more alleles is downregulatedto produce the upright stature phenotype. In some embodiments, theupright stature phenotype is the result of gene expression andenvironmental conditions (e.g., heat). In some embodiments, the uprightstature phenotype is the result of gene expression and cultivationconditions (e.g., close planting). In some embodiments, specificcharacteristics of upright stature can be linked to specific alleles(e.g., increased height to height allele at locus B). In someembodiments, linked markers can be used to identify these alleles.

Certain aspects of the present disclosure relate to an iceberg lettuceseed from the plants of any of the above embodiments. In anotherembodiment, the present disclosure is directed to iceberg lettuce plantsgrown from the seed of any of the above embodiments. In some embodimentsof any of the above embodiments, the iceberg lettuce plant includes aspace between a base of a head and a top of a ground, and said spaceincludes an increased length of core outside of a processing material ofa head (i.e., external stem). In some embodiments, the increased lengthof core outside of the processing material of the head (i.e., externalstem) is about 3.5 cm or more, about 4 cm or more, about 4.5 cm or more,or about 5 cm or more. In some embodiments, the iceberg lettuce plantfurther includes one or more characteristics from the group of: a heightto diameter ratio of about 1.3 to about 1.5, leaf strength between about300 to 400 grams, about 40 to 50% overlapping leaves inside theprocessing material of the head, a short stem inside of the processingmaterial of the head (i.e., internal stem), leaves with about 4.8 to5.3° Brix, leaves with about 25 to 40 g/l total sugars, leaves withabout 4.5 to 5% dry matter weight, 5-10% processing damage per bag ofprocessed plant material, and a range of about 35 to 42 days of shelflife of processed and packaged plant material.

In another embodiment, the present disclosure is directed to methods ofproducing an herbicide resistant lettuce plant by introducing a geneconferring herbicide resistance, where the gene is selected fromglyphosate, sulfonylurea, imidazolinone, dicamba, glufosinate, phenoxyproprionic acid, L-phosphinothricin, cyclohexone, cyclohexanedione,triazine, and benzonitrile, into a lettuce plant produced by growinglettuce seed of any of the above embodiments, and to herbicide resistantlettuce plants produced by such methods. In another embodiment, thepresent disclosure is directed to methods of producing a pest or insectresistant lettuce plan by introducing a gene conferring pest or insectresistance (e.g., a gene encoding a Bacillus thuringiensis endotoxin)into a lettuce plant produced by growing lettuce seed of any of theabove embodiments, and to pest or insect resistant lettuce plantsproduced by such methods. In another embodiment, the present disclosureis directed to methods of producing a disease resistant lettuce plant byintroducing a gene conferring disease resistance into a lettuce plantproduced by growing lettuce seed of any of the above embodiments, and todisease resistant lettuce plants produced by such methods. In anotherembodiment, the present disclosure is directed to methods of producing alettuce plant with a value-added trait by introducing a gene conferringa value-added trait, where the gene encodes a protein selected from aferritin, a nitrate reductase, and a monellin into a lettuce plantproduced by growing lettuce seed of any of the above embodiments, and tovalue-added lettuce plants produced by such methods. In anotherembodiment, the present disclosure is directed to methods of producing alettuce plant adapted to specific environmental conditions (e.g.,drought, flooding, salinity, low nitrogen, high heat, high CO₂, etc.),into a lettuce plant produced by growing lettuce seed of any of theabove embodiments, and to lettuce plants adapted to specificenvironmental conditions produced by such methods. In anotherembodiment, the present disclosure provides for single gene convertedplants of any of the above embodiments. The single transferred gene maypreferably be a dominant or recessive allele. Preferably, the singletransferred gene will confer such traits as male sterility, herbicideresistance, and insect or pest resistance, modified fatty acidmetabolism, modified carbohydrate metabolism, resistance for bacterial,fungal, or viral disease, male fertility, enhanced nutritional quality,and industrial usage. The single gene may be a naturally occurringlettuce gene.

Methods of Generating and Selecting Upright Heading Iceberg LettucePlants

Certain aspects of the present disclosure relate to methods ofgenerating and selecting upright heading iceberg lettuce plants, and theseeds that produce the upright heading iceberg lettuce plants describedherein.

In some embodiments, the method of generating and selecting uprightheading iceberg lettuce plants involved crossing a first parentaliceberg lettuce plant and a second parental iceberg lettuce plant. Insome embodiments, the first parental iceberg lettuce plant is the maleparental iceberg lettuce plant and the second parental iceberg lettuceplant is the female parental iceberg lettuce plant. In some embodiments,the first parental iceberg lettuce plant is the female parental iceberglettuce plant and the second parental iceberg lettuce plant is the maleparental iceberg lettuce plant. In some embodiments, the first parentaliceberg lettuce plant and the second iceberg lettuce plant are plantsfrom inbred iceberg lettuce plant lines. In some embodiments, the firstparental iceberg lettuce plant and the second iceberg lettuce plant areoff-types (e.g., rogues) from inbred iceberg lettuce plant lines. Insome embodiments, the iceberg lettuce plant lines are the same. In otherembodiments, the iceberg lettuce plant lines are different. In someembodiments, one of the parental iceberg lettuce plants is a plant ofany of the above embodiments and the other parental iceberg lettuceplant is another iceberg lettuce plant. In some embodiments, both of theparental iceberg lettuce plants are a plant of any of the aboveembodiments. In some embodiments, one of the parental iceberg lettuceplants is ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, or‘E01E.70168’ Lot B. In some embodiments, one of the parental iceberglettuce plants and the other parental iceberg lettuce plant are fromdifferent lines (e.g., ‘E01E.70111‘ Lot A and’E01E.70111’ Lot B). Inanother embodiment, one of the parental iceberg lettuce plants and theother parental iceberg lettuce plant are from the same line (e.g.,‘E01E.70111’ Lot A). In still another embodiment, the present disclosureis further directed to lettuce plants, lettuce parts from the lettuceplants, and seeds produced therefrom where the lettuce plant is isolatedby the breeding method of the disclosure.

In another embodiment, the present disclosure is further directed to amethod of selecting lettuce plants with an upright stature, by a)crossing a first lettuce plant with a height to diameter ratio greaterthan or equal to about 1.0, a height of about 1.5 to about 3 times aheight of a standard iceberg lettuce plant, a crisp leaf texture, and aclosed head with a second lettuce plant with a height to diameter ratiogreater than or equal to about 1.0, a height of about 1.5 to about 3times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head, b) selecting offspring with an improved setof characteristics including a height to diameter ratio greater than orequal to about 1.5, a height greater than or equal to about 2 to about 3times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a more tightly closed head, c) selfing or sib-crossing(i.e. sobbing) the offspring, and d) repeating steps b) and c) formultiple generations to produce inbred lines with the improved set ofcharacteristics. In one embodiment, the characteristics used forselection in step b) further include one or more characteristics fromthe group of: a short core, a low level of internal tipburn, a low levelof bottom rot, an increased number of leaves, a fast fill rate of leavesin a head, and a space between a base of a head and a top of a ground.In another embodiment, the space between the base of the head and thetop of the ground is achieved by an increased length of core or anincreased number of frame leaves outside of the processing material ofthe head. In some embodiments, the characteristics used for selection instep b) further include one or more characteristics from the group of:moderate leaf glossiness, moderate tendency to bolt, inner leaves palerthan outer leaves, blanched inner leaves, green to greyish green outerleaves, outer leaves with a color ranging from about RHS 146A to aboutRHS146B, inner leaves with a color ranging from RHS 145C to RHS 245D,leaves with an obovate shape, leaves with an elliptic shape, leaves witha broad obtrullate shape, leaves with a triangular shape, head with anarrow elliptic shape in longitudinal section, head with an ellipticshape in longitudinal section, head with a broad elliptic shape inlongitudinal section, head with an oval shape in longitudinal section,head with an oblong shape in longitudinal section, medium degree ofoverlapping of upper part of leaves, strong degree of overlapping ofupper part of leaves, absent or very few leaf divisions, leaf marginhardly to rather strongly incised, thick leaves, absent to mediumundulation of leaf margin, semi-flabellate leaf venation, flabellateleaf venation, clear midrib, no clear midrib, heading, early heading,and medium heading. In other embodiments, the iceberg lettuce plantfurther includes one or more characteristics from the group ofcharacteristics defining the upright heading iceberg lettuce plant type.In still another embodiment, the present disclosure is further directedto lettuce plants, lettuce parts from the lettuce plants, and seedsproduced therefrom where the lettuce plant is isolated by the breedingmethod of the disclosure.

In another embodiment, the present disclosure is further directed to amethod of making upright heading iceberg lettuce plants, by a) crossinga first upright heading iceberg lettuce plant including a space betweena base of a head and a top of a ground, wherein said space includes anincreased length of core outside of a processing material of a head(i.e., external stem) with a second upright heading iceberg lettuceplant including a space between a base of a head and a top of a ground,wherein said space includes an increased length of core outside of aprocessing material of a head (i.e., external stem); b) selectingoffspring lettuce plants including a space between a base of a head anda top of a ground, wherein said space includes an increased length ofcore outside of a processing material of a head (i.e., external stem);c) selfing or sibbing the offspring, and d) repeating steps b) and c)for multiple generations to produce inbred lines with the improved setof characteristics. In some embodiments, the increased length of coreoutside of the processing material of the head (i.e., external stem) isabout 3.5 cm or more, about 4 cm or more, about 4.5 cm or more, or about5 cm or more. In one embodiment, the characteristics used for selectionin step b) further include one or more characteristics from the groupof: a height to diameter ratio of about 1.3 to about 1.5, leaf strengthbetween about 300 to 400 grams, about 40 to 50% overlapping leavesinside the processing material of the head, a short stem inside of theprocessing material of the head (i.e., internal stem), leaves with about4.8 to 5.3° Brix, leaves with about 25 to 40 g/l total sugars, leaveswith about 4.5 to 5% dry matter weight, 5-10% processing damage per bagof processed plant material, and a range of about 35 to 42 days of shelflife of processed and packaged plant material.

In a further embodiment, the present disclosure relates to methods fordeveloping lettuce plants in a lettuce plant breeding program usingplant breeding techniques including recurrent selection, backcrossing,pedigree breeding, restriction fragment length polymorphism enhancedselection, genetic marker enhanced selection, and transformation. Seeds,lettuce plants, and parts thereof, produced by such breeding methods arealso part of the disclosure.

Further Embodiments

Gene Conversions

When the term “lettuce plant” is used in the context of the presentinvention, this also includes any gene conversions of that variety. Theterm “gene converted plant” as used herein refers to those lettuceplants which are developed by backcrossing, genetic engineering, ormutation, where essentially all of the desired morphological andphysiological characteristics of a variety are recovered in addition tothe one or more genes transferred into the variety via the backcrossingtechnique, genetic engineering, or mutation. Backcrossing methods can beused with the present invention to improve or introduce a characteristicinto the variety. The term “backcrossing” as used herein refers to therepeated crossing of a hybrid progeny back to the recurrent parent,i.e., backcrossing 1, 2, 3, 4, 5, 6, 7, 8, 9, or more times to therecurrent parent. The parental lettuce plant which contributes the genefor the desired characteristic is termed the “nonrecurrent” or “donorparent.” This terminology refers to the fact that the nonrecurrentparent is used one time in the backcross protocol and therefore does notrecur. The parental lettuce plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol. Perlman &Sleeper (1994) and Fehr (1993). In a typical backcross protocol, theoriginal variety of interest (recurrent parent) is crossed to a secondvariety (nonrecurrent parent) that carries the gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until alettuce plant is obtained where essentially all of the desiredmorphological and physiological characteristics of the recurrent parentare recovered in the converted plant, in addition to the transferredgene from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a trait or characteristic in the original line.To accomplish this, a gene of the recurrent variety is modified orsubstituted with the desired gene from the nonrecurrent parent, whileretaining essentially all of the rest of the desired genetic, andtherefore the desired physiological and morphological, constitution ofthe original line. The choice of the particular nonrecurrent parent willdepend on the purpose of the backcross. One of the major purposes is toadd some commercially desirable, agronomically important trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered to determine an appropriate testing protocol.Although backcrossing methods are simplified when the characteristicbeing transferred is a dominant allele, a recessive allele may also betransferred. In this instance it may be necessary to introduce a test ofthe progeny to determine if the desired characteristic has beensuccessfully transferred.

Many gene traits have been identified that are not regularly selected inthe development of a new line but that can be improved by backcrossingtechniques. Examples of these traits include, but are not limited to,male sterility, modified fatty acid metabolism, modified carbohydratemetabolism, herbicide resistance, resistance for bacterial, fungal, orviral disease, insect resistance, enhanced nutritional quality,industrial usage, yield stability, and yield enhancement. These genesare generally inherited through the nucleus. Several of these genetraits are described in U.S. Pat. Nos. 5,777,196, 5,948,957, and5,969,212, the disclosures of which are specifically hereby incorporatedby reference.

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of lettuce andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to Tang, et al., HortScience, 27:9,1030-1032 (1992); Teng, et al., HortScience, 28:6, 669-1671 (1993);Zhang, et al., Journal of Genetics and Breeding, 46:3, 287-290 (1992);Webb, et al., Plant Cell Tissue and Organ Culture, 38:1, 77-79 (1994);Curtis, et al., Journal of Experimental Botany, 45:279, 1441-1449(1994); Nagata, et al., Journal for the American Society forHorticultural Science, 125:6, 669-672 (2000); and Ibrahim, et al., PlantCell Tissue and Organ Culture, 28(2), 139-145 (1992). It is clear fromthe literature that the state of the art is such that these methods ofobtaining plants are routinely used and have a very high rate ofsuccess. Thus, another aspect of this invention is to provide cellswhich upon growth and differentiation produce lettuce plants having thephysiological and morphological characteristics of variety ‘E01E.70111’.A further aspect of this invention is to provide cells which upon growthand differentiation produce lettuce plants having the physiological andmorphological characteristics of variety ‘E01E.70168’.

As used herein, the term “tissue culture” indicates a compositioncontaining isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, suckers, and the like. Meansfor preparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture containing organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and5,977,445 describe certain techniques, the disclosures of which areincorporated herein by reference.

Additional Breeding Methods

The invention is also directed to methods for producing a lettuce plantby crossing a first parent lettuce plant with a second parent lettuceplant where the first or second parent lettuce plant is a lettuce plantof variety ‘E01E.70111’ Lot A. Further, both first and second parentlettuce plants can come from lettuce variety ‘E01E.70111’ Lot A. Thus,any such methods using lettuce variety ‘E01E.70111’ Lot A are part ofthe invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using lettuce variety‘E01E.70111’ Lot A as at least one parent are within the scope of thisinvention, including those developed from varieties derived from lettucevariety ‘E01E.70111’ Lot A. Advantageously, this lettuce variety couldbe used in crosses with other, different, lettuce plants to produce thefirst generation (F₁) lettuce hybrid seeds and plants with superiorcharacteristics. The variety of the invention can also be used fortransformation where exogenous genes are introduced and expressed by thevariety of the invention. Genetic variants created either throughtraditional breeding methods using lettuce variety ‘E01E.70111’ Lot A,or through transformation of variety ‘E01E.70111’ Lot A by any of anumber of protocols known to those of skill in the art are intended tobe within the scope of this invention.

The following describes breeding methods that may be used with lettucevariety ‘E01E.70111’ Lot A in the development of further lettuce plants.One such embodiment is a method for developing variety ‘E01E.70111’ LotA progeny lettuce plants in a lettuce plant breeding program, by:obtaining the lettuce plant, or a part thereof, of variety ‘E01E.70111’Lot A, utilizing said plant or plant part as a source of breedingmaterial, and selecting a lettuce variety ‘E01E.70111’ Lot A progenyplant with molecular markers in common with variety ‘E01E.70111’ Lot Aand/or with morphological and/or physiological characteristics selectedfrom the characteristics listed in the section entitled “Objectivedescription of the variety ‘E01E.70111’ Lot A” Breeding steps that maybe used in the lettuce plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers), and the making ofdouble haploids may be utilized.

Another method involves producing a population of lettuce variety‘E01E.70111’ Lot A progeny lettuce plants, by crossing variety‘E01E.70111’ Lot A with another lettuce plant, thereby producing apopulation of lettuce plants, which, on average, derive 50% of theiralleles from lettuce variety ‘E01E.70111’ Lot A. A plant of thispopulation may be selected and repeatedly selfed or sibbed with alettuce variety resulting from these successive filial generations. Oneembodiment of this invention is the lettuce variety produced by thismethod and that has obtained at least 50% of its alleles from lettucevariety ‘E01E.70111’ Lot A. One of ordinary skill in the art of plantbreeding would know how to evaluate the traits of two plant varieties todetermine if there is no significant difference between the two traitsexpressed by those varieties. For example, see Fehr and Walt, Principlesof Variety Development, pp. 261-286 (1987). Thus the invention includeslettuce variety ‘E01E.70111’ Lot A progeny lettuce plants containing acombination of at least two variety ‘E01E.70111’ Lot A traits selectedfrom those listed in the section entitled “Objective description of thevariety ‘E01E.70111’ Lot A”, or the variety ‘E01E.70111’ Lot Acombination of traits listed in the Summary of the Invention, so thatsaid progeny lettuce plant is not significantly different for saidtraits than lettuce variety ‘E01E.70111’ Lot A as determined at the 5%significance level when grown in the same environmental conditions.Using techniques described herein, molecular markers may be used toidentify said progeny plant as a lettuce variety ‘E01E.70111’ Lot Aprogeny plant. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions. Once such avariety is developed, its value is substantial since it is important toadvance the germplasm base as a whole in order to maintain or improvetraits such as yield, disease resistance, pest resistance, and plantperformance in extreme environmental conditions.

Progeny of lettuce variety ‘E01E.70111’ Lot A may also be characterizedthrough their filial relationship with lettuce variety ‘E01E.70111’ LotA, as for example, being within a certain number of breeding crosses oflettuce variety ‘E01E.70111’ Lot A. A breeding cross is a cross made tointroduce new genetics into the progeny, and is distinguished from across, such as a self or a sib cross, made to select among existinggenetic alleles. The lower the number of breeding crosses in thepedigree, the closer the relationship between lettuce variety‘E01E.70111’ Lot A and its progeny. For example, progeny produced by themethods described herein may be within 1, 2, 3, 4, or 5 breeding crossesof lettuce variety ‘E01E.70111’ Lot A.

The invention is also directed to methods for producing a lettuce plantby crossing a first parent lettuce plant with a second parent lettuceplant where the first or second parent lettuce plant is a lettuce plantof variety ‘E01E.70111’ Lot B. Further, both first and second parentlettuce plants can come from lettuce variety ‘E01E.70111’ Lot B. Thus,any such methods using lettuce variety ‘E01E.70111’ Lot B are part ofthe invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using lettuce variety‘E01E.70111’ Lot B as at least one parent are within the scope of thisinvention, including those developed from varieties derived from lettucevariety ‘E01E.70111’ Lot B. Advantageously, this lettuce variety couldbe used in crosses with other, different, lettuce plants to produce thefirst generation (F₁) lettuce hybrid seeds and plants with superiorcharacteristics. The variety of the invention can also be used fortransformation where exogenous genes are introduced and expressed by thevariety of the invention. Genetic variants created either throughtraditional breeding methods using lettuce variety ‘E01E.70111’ Lot B,or through transformation of variety ‘E01E.70111’ Lot B by any of anumber of protocols known to those of skill in the art are intended tobe within the scope of this invention.

The following describes breeding methods that may be used with lettucevariety ‘E01E.70111’ Lot B in the development of further lettuce plants.One such embodiment is a method for developing variety ‘E01E.70111’ LotB progeny lettuce plants in a lettuce plant breeding program, by:obtaining the lettuce plant, or a part thereof, of variety ‘E01E.70111’Lot B, utilizing said plant or plant part as a source of breedingmaterial, and selecting a lettuce variety ‘E01E.70111’ Lot B progenyplant with molecular markers in common with variety ‘E01E.70111’ Lot Band/or with morphological and/or physiological characteristics selectedfrom the characteristics listed in the section entitled “Objectivedescription of the variety ‘E01E.70111’ Lot B” Breeding steps that maybe used in the lettuce plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers), and the making ofdouble haploids may be utilized.

Another method involves producing a population of lettuce variety‘E01E.70111’ Lot B progeny lettuce plants, by crossing variety‘E01E.70111’ Lot B with another lettuce plant, thereby producing apopulation of lettuce plants, which, on average, derive 50% of theiralleles from lettuce variety ‘E01E.70111’ Lot B. A plant of thispopulation may be selected and repeatedly selfed or sibbed with alettuce variety resulting from these successive filial generations. Oneembodiment of this invention is the lettuce variety produced by thismethod and that has obtained at least 50% of its alleles from lettucevariety ‘E01E.70111’ Lot B. One of ordinary skill in the art of plantbreeding would know how to evaluate the traits of two plant varieties todetermine if there is no significant difference between the two traitsexpressed by those varieties. For example, see Fehr and Walt, Principlesof Variety Development, pp. 261-286 (1987). Thus the invention includeslettuce variety ‘E01E.70111’ Lot B progeny lettuce plants containing acombination of at least two variety ‘E01E.70111’ Lot B traits selectedfrom those listed in the section entitled “Objective description of thevariety ‘E01E.70111’ Lot B”, or the variety ‘E01E.70111’ Lot Bcombination of traits listed in the Summary of the Invention, so thatsaid progeny lettuce plant is not significantly different for saidtraits than lettuce variety ‘E01E.70111’ Lot B as determined at the 5%significance level when grown in the same environmental conditions.Using techniques described herein, molecular markers may be used toidentify said progeny plant as a lettuce variety ‘E01E.70111’ Lot Bprogeny plant. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions. Once such avariety is developed, its value is substantial since it is important toadvance the germplasm base as a whole in order to maintain or improvetraits such as yield, disease resistance, pest resistance, and plantperformance in extreme environmental conditions.

Progeny of lettuce variety ‘E01E.70111’ Lot B may also be characterizedthrough their filial relationship with lettuce variety ‘E01E.70111’ LotB, as for example, being within a certain number of breeding crosses oflettuce variety ‘E01E.70111’ Lot B. A breeding cross is a cross made tointroduce new genetics into the progeny, and is distinguished from across, such as a self or a sib cross, made to select among existinggenetic alleles. The lower the number of breeding crosses in thepedigree, the closer the relationship between lettuce variety‘E01E.70111’ Lot B and its progeny. For example, progeny produced by themethods described herein may be within 1, 2, 3, 4, or 5 breeding crossesof lettuce variety ‘E01E.70111’ Lot B.

The invention is also directed to methods for producing a lettuce plantby crossing a first parent lettuce plant with a second parent lettuceplant where the first or second parent lettuce plant is a lettuce plantof variety ‘E01E.70168’ Lot A. Further, both first and second parentlettuce plants can come from lettuce variety “E01E.70168’ Lot A’. Thus,any such methods using lettuce variety ‘E01E.70168’ Lot A are part ofthe invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using lettuce variety‘E01E.70168’ Lot A as at least one parent are within the scope of thisinvention, including those developed from varieties derived from lettucevariety ‘E01E.70168’ Lot A. Advantageously, this lettuce variety couldbe used in crosses with other, different, lettuce plants to produce thefirst generation (F₁) lettuce hybrid seeds and plants with superiorcharacteristics. The variety of the invention can also be used fortransformation where exogenous genes are introduced and expressed by thevariety of the invention. Genetic variants created either throughtraditional breeding methods using lettuce variety ‘E01E.70168’ Lot A,or through transformation of variety ‘E01E.70168’ Lot A by any of anumber of protocols known to those of skill in the art are intended tobe within the scope of this invention.

The following describes breeding methods that may be used with lettucevariety ‘E01E.70168’ Lot A in the development of further lettuce plants.One such embodiment is a method for developing variety ‘E01E.70168’ LotA progeny lettuce plants in a lettuce plant breeding program, by:obtaining the lettuce plant, or a part thereof, of variety ‘E01E.70168’Lot A, utilizing said plant or plant part as a source of breedingmaterial, and selecting a lettuce variety ‘E01E.70168’ Lot A progenyplant with molecular markers in common with variety ‘E01E.70168’ Lot Aand/or with morphological and/or physiological characteristics selectedfrom the characteristics listed in the section entitled “Objectivedescription of the variety ‘E01E.70168’ Lot A”. Breeding steps that maybe used in the lettuce plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers), and the making ofdouble haploids may be utilized.

Another method involves producing a population of lettuce variety‘E01E.70168’ Lot A progeny lettuce plants, by crossing variety‘E01E.70168’ Lot A with another lettuce plant, thereby producing apopulation of lettuce plants, which, on average, derive 50% of theiralleles from lettuce variety ‘E01E.70168’ Lot A. A plant of thispopulation may be selected and repeatedly selfed or sibbed with alettuce variety resulting from these successive filial generations. Oneembodiment of this invention is the lettuce variety produced by thismethod and that has obtained at least 50% of its alleles from lettucevariety ‘E01E.70168’ Lot A. One of ordinary skill in the art of plantbreeding would know how to evaluate the traits of two plant varieties todetermine if there is no significant difference between the two traitsexpressed by those varieties. For example, see Fehr and Walt, Principlesof Variety Development, pp. 261-286 (1987). Thus the invention includeslettuce variety ‘E01E.70168’ Lot A progeny lettuce plants containing acombination of at least two variety ‘E01E.70168’ Lot A traits selectedfrom those listed in the section entitled “Objective description of thevariety ‘E01E.70168’ Lot A”, or the variety ‘E01E.70168’ Lot Acombination of traits listed in the Summary of the Invention, so thatsaid progeny lettuce plant is not significantly different for saidtraits than lettuce variety ‘E01E.70168’ Lot A as determined at the 5%significance level when grown in the same environmental conditions.Using techniques described herein, molecular markers may be used toidentify said progeny plant as a lettuce variety ‘E01E.70168’ Lot Aprogeny plant. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions. Once such avariety is developed, its value is substantial since it is important toadvance the germplasm base as a whole in order to maintain or improvetraits such as yield, disease resistance, pest resistance, and plantperformance in extreme environmental conditions.

Progeny of lettuce variety ‘E01E.70168’ Lot A may also be characterizedthrough their filial relationship with lettuce variety ‘E01E.70168’ LotA, as for example, being within a certain number of breeding crosses oflettuce variety ‘E01E.70168’ Lot A. A breeding cross is a cross made tointroduce new genetics into the progeny, and is distinguished from across, such as a self or a sib cross, made to select among existinggenetic alleles. The lower the number of breeding crosses in thepedigree, the closer the relationship between lettuce variety‘E01E.70168’ Lot A and its progeny. For example, progeny produced by themethods described herein may be within 1, 2, 3, 4, or 5 breeding crossesof lettuce variety ‘E01E.70168’ Lot A.

The invention is also directed to methods for producing a lettuce plantby crossing a first parent lettuce plant with a second parent lettuceplant where the first or second parent lettuce plant is a lettuce plantof variety ‘E01E.70168’ Lot B. Further, both first and second parentlettuce plants can come from lettuce variety “E01E.70168’ Lot B’. Thus,any such methods using lettuce variety ‘E01E.70168’ Lot B are part ofthe invention: selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using lettuce variety‘E01E.70168’ Lot B as at least one parent are within the scope of thisinvention, including those developed from varieties derived from lettucevariety ‘E01E.70168’ Lot B. Advantageously, this lettuce variety couldbe used in crosses with other, different, lettuce plants to produce thefirst generation (F₁) lettuce hybrid seeds and plants with superiorcharacteristics. The variety of the invention can also be used fortransformation where exogenous genes are introduced and expressed by thevariety of the invention. Genetic variants created either throughtraditional breeding methods using lettuce variety ‘E01E.70168’ Lot B,or through transformation of variety ‘E01E.70168’ Lot B by any of anumber of protocols known to those of skill in the art are intended tobe within the scope of this invention.

The following describes breeding methods that may be used with lettucevariety ‘E01E.70168’ Lot B in the development of further lettuce plants.One such embodiment is a method for developing variety ‘E01E.70168’ LotB progeny lettuce plants in a lettuce plant breeding program, by:obtaining the lettuce plant, or a part thereof, of variety ‘E01E.70168’Lot B, utilizing said plant or plant part as a source of breedingmaterial, and selecting a lettuce variety ‘E01E.70168’ Lot B progenyplant with molecular markers in common with variety ‘E01E.70168’ Lot Band/or with morphological and/or physiological characteristics selectedfrom the characteristics listed in the section entitled “Objectivedescription of the variety ‘E01E.70168’ Lot B”. Breeding steps that maybe used in the lettuce plant breeding program include pedigree breeding,backcrossing, mutation breeding, and recurrent selection. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers), and the making ofdouble haploids may be utilized.

Another method involves producing a population of lettuce variety‘E01E.70168’ Lot B progeny lettuce plants, by crossing variety‘E01E.70168’ Lot B with another lettuce plant, thereby producing apopulation of lettuce plants, which, on average, derive 50% of theiralleles from lettuce variety ‘E01E.70168’ Lot B. A plant of thispopulation may be selected and repeatedly selfed or sibbed with alettuce variety resulting from these successive filial generations. Oneembodiment of this invention is the lettuce variety produced by thismethod and that has obtained at least 50% of its alleles from lettucevariety ‘E01E.70168’ Lot B. One of ordinary skill in the art of plantbreeding would know how to evaluate the traits of two plant varieties todetermine if there is no significant difference between the two traitsexpressed by those varieties. For example, see Fehr and Walt, Principlesof Variety Development, pp. 261-286 (1987). Thus the invention includeslettuce variety ‘E01E.70168’ Lot B progeny lettuce plants containing acombination of at least two variety ‘E01E.70168’ Lot B traits selectedfrom those listed in the section entitled “Objective description of thevariety ‘E01E.70168’ Lot B”, or the variety ‘E01E.70168’ Lot Bcombination of traits listed in the Summary of the Invention, so thatsaid progeny lettuce plant is not significantly different for saidtraits than lettuce variety ‘E01E.70168’ Lot B as determined at the 5%significance level when grown in the same environmental conditions.Using techniques described herein, molecular markers may be used toidentify said progeny plant as a lettuce variety ‘E01E.70168’ Lot Bprogeny plant. Mean trait values may be used to determine whether traitdifferences are significant, and preferably the traits are measured onplants grown under the same environmental conditions. Once such avariety is developed, its value is substantial since it is important toadvance the germplasm base as a whole in order to maintain or improvetraits such as yield, disease resistance, pest resistance, and plantperformance in extreme environmental conditions.

Progeny of lettuce variety ‘E01E.70168’ Lot B may also be characterizedthrough their filial relationship with lettuce variety ‘E01E.70168’ LotB, as for example, being within a certain number of breeding crosses oflettuce variety ‘E01E.70168’ Lot B. A breeding cross is a cross made tointroduce new genetics into the progeny, and is distinguished from across, such as a self or a sib cross, made to select among existinggenetic alleles. The lower the number of breeding crosses in thepedigree, the closer the relationship between lettuce variety‘E01E.70168’ Lot B and its progeny. For example, progeny produced by themethods described herein may be within 1, 2, 3, 4, or 5 breeding crossesof lettuce variety ‘E01E.70168’ Lot B.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which lettuce plants can beregenerated, plant calli, plant clumps, and plant cells that are intactin plants or parts of plants, such as leaves, pollen, embryos,cotyledons, hypocotyl, roots, root tips, anthers, pistils, flowers,ovules, seeds, stems, and the like.

The use of the terms “a,” “an,” and “the,” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Forexample, if the range 10-15 is disclosed, then 11, 12, 13, and 14 arealso disclosed. All methods described herein can be performed in anysuitable order unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the invention and does not pose a limitation on the scope ofthe invention unless otherwise claimed. No language in the specificationshould be construed as indicating any non-claimed element as essentialto the practice of the invention.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions, and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions, and sub-combinations as are within their truespirit and scope.

Overview of the Variety ‘E01E.70111’ Lot A

‘E01E.70111’ Lot A is an inbred line from an original cross between twoiceberg rogues. ‘E01E.70111‘ and’E01E.70168’ are inbred lines from thesame original cross. ‘E01E.70111‘ Lot A and’E01E.70111’ Lot B are sisterlines from the same parent in the previous generation.

Lettuce variety ‘E01E.70111’ Lot A is the result of numerous generationsof plant selections chosen for its oblong to oval shape, taller height,crisp leaf texture, and closed head. Further traits of interest includedits tighter filling head, short core, lower level of internal tip-burn,lower level of bottom rot, and the elevation of its base from theground.

The variety has shown uniformity and stability for the traits, withinthe limits of environmental influence for the traits. It has beenself-pollinated a sufficient number of generations with carefulattention to uniformity of plant type. The line has been increased withcontinued observation for uniformity. No variant traits have beenobserved or are expected in variety ‘E01E.70111’ Lot A.

The data which define these characteristics is based on observationstaken in San Juan Bautista, Calif. Color references are primarily to theRHS Colour Chart of The Royal Horticultural Society of London (RHS)(2007 edition). Descriptive terminology follows the Plant IdentificationTerminology, An Illustrated Glossary, 2nd edition by James G. Harris andMelinda Woolf Harris, unless where otherwise defined.

Objective Description of the Variety ‘E01E.70111’ Lot A

Lettuce variety ‘E01E.70111’ Lot A has the following morphologic andother characteristics:

Plant:

-   -   Type: Upright heading iceberg    -   Shape: Oblong to oval    -   Height: Tall    -   Diameter: Narrow    -   Core length: Relatively short under cool conditions; long cores        under high heat conditions

Leaves:

-   -   Heading:        -   Outer leaves: Curl over, less tight wrap than iceberg        -   Inner leaves: Blanched    -   Color:        -   Outer leaves: Medium green        -   Inner leaves: Yellow-green    -   Mature leaf texture: Crisp        Comparisons to Other Lettuce Varieties

Table 1 below compares the characteristics of lettuce variety‘E01E.70111’ Lot A with the most similar varieties ‘Pueblo’ (icebergtype, U.S. patent application Ser. No. 15/448,038), ‘True Heart’(romaine type, U.S. Pat. No. 7,468,473), and ‘Crunchita’ (cosberg type,U.S. Pat. No. 9,119,366). Table 2 below compares the qualities of theharvested material of these varieties, including shelf life, maturityindex, and other qualities.

TABLE 1 ‘E01E.70111’ Characteristic Lot A ‘Pueblo’ ‘True Heart’‘Crunchita’ Head shape Oval to oblong Round V-shaped/ Oval to oblongDeep V-shaped Leaf shape Fan-shaped to Fan-shaped V-shaped/ Fan-shapedto V-shaped/ Deep V-shaped V-shaped/ Deep V-shaped Deep V-shaped (moreFan-shaped) (more V-shaped)

TABLE 2 ‘E01E.70111’ Characteristic Lot A ‘Pueblo’ ‘True Heart’‘Crunchita’ Shelf life (days post harvest) 22 22 16 16 Odor (days postharvest) 22 22 14 14 Browning (days post harvest) 22 22 16 16 Rotting(days post harvest) 20 20 14 14 Maturity index (1 to 9) 2.4 ± 0.5 4.1 ±0.7 38.8 ± 17.7 36.8 ± 12.6 Processing efficiency (%) 82.3 ± 2.1  91.6 ±4.1  84.7 ± 3.3  89.7 ± 2.7  Internal tipburn (1 to 9) 8.3 7.3 7.8 8.6Bremia (1 to 9) 9.0 7.7 9.0 9.0 Splitting (1 to 9) 7.5 4.3 6.0 8.6 Sideshoot (1 to 9) 9.0 9.0 9.0 9.0Overview of the Variety ‘E01E.70111’ Lot B

‘E01E.70111’ Lot B is an inbred line from an original cross between twoiceberg rogues. ‘E01E.70111‘ and’E01E.70168’ are inbred lines from thesame original cross. ‘E01E.70111‘ Lot A and’E01E.70111’ Lot B are sisterlines from the same parent in the previous generation.

Lettuce variety ‘E01E.70111’ Lot B is the result of numerous generationsof plant selections chosen for its oblong to oval shape, taller height,crisp leaf texture, and closed head. Further traits of interest includedits tighter filling head, short core, lower level of internal tip-burn,lower level of bottom rot, and the elevation of its base from theground.

The variety has shown uniformity and stability for the traits, withinthe limits of environmental influence for the traits. It has beenself-pollinated a sufficient number of generations with carefulattention to uniformity of plant type. The line has been increased withcontinued observation for uniformity. No variant traits have beenobserved or are expected in variety ‘E01E.70111’ Lot B.

The data which define these characteristics is based on observationstaken in San Juan Bautista, Calif. Color references are primarily to theRHS Colour Chart of The Royal Horticultural Society of London (RHS)(2007 edition). Descriptive terminology follows the Plant IdentificationTerminology, An Illustrated Glossary, 2nd edition by James G. Harris andMelinda Woolf Harris, unless where otherwise defined.

Objective Description of the Variety ‘E01E.70111’ Lot B

Lettuce variety ‘E01E.70111’ Lot B has the following morphologic andother characteristics:

Plant:

-   -   Type: Upright heading iceberg    -   Shape: Oblong to oval    -   Height: Tall    -   Diameter: Narrow    -   Core length: Relatively short under cool conditions; long cores        under high heat conditions

Leaves:

-   -   Heading:        -   Outer leaves: Curl over, less tight wrap than iceberg        -   Inner leaves: Blanched    -   Color:        -   Outer leaves: Medium green        -   Inner leaves: Yellow-green    -   Mature leaf texture: Crisp        Comparisons to Other Lettuce Varieties

Table 3 below compares the characteristics of lettuce variety‘E01E.70111’ Lot B with the most similar varieties ‘Pueblo’ (icebergtype), ‘True Heart’ (romaine type), and ‘Crunchita’ (cosberg type).Table 4 below compares the qualities of the harvested material of thesevarieties, including shelf life, maturity index, and other qualities.

TABLE 3 ‘E01E.70111’ ‘True Characteristic Lot B ‘Pueblo’ Heart’‘Crunchita’ Head shape Oval to oblong Round V-shaped/ Oval to oblongDeep V-shaped Leaf shape Fan-shaped to Fan- V-shaped/ Fan-shaped toV-shaped/ shaped Deep V-shaped/ Deep V-shaped Deep V-shaped V-shaped(more (more V-shaped) Fan-shaped)

TABLE 4 ‘E01E.70111’ ‘True Characteristic Lot B ‘Pueblo’ Heart’‘Crunchita’ Shelf life (days post 20 22 16 16 harvest) Odor (days postharvest) 20 22 14 14 Browning (days post 18 22 16 16 harvest) Rotting(days post 18 20 14 14 harvest) Maturity index (1 to 9) 1.8 ± 0.7 4.1 ±0.7 38.8 ± 17.7 36.8 ± 12.6 Processing efficiency (%) 81.1 ± 2.2  91.6 ±4.1  84.7 ± 3.3  89.7 ± 2.7  Internal tipburn (1 to 9) 6.8 7.3 7.8 8.6Bremia (1 to 9) 9.0 7.7 9.0 9.0 Splitting (1 to 9) 7.1 4.3 6.0 8.6 Sideshoot (1 to 9) 9.0 9.0 9.0 9.0Overview of the Variety ‘E01E.70168’ Lot A

‘E01E.70168’ Lot A is an inbred line from an original cross between twoiceberg rogues. ‘E01E.70111‘ and’E01E.70168’ are inbred lines from thesame original cross. ‘E01E.70168‘ Lot A and’E01E.70168’ Lot B are sisterlines from the same parent in the previous generation.

Lettuce variety ‘E01E.70168’ Lot A is the result of numerous generationsof plant selections chosen for its oblong to oval shape, taller height,crisp leaf texture, and closed head. Further traits of interest includedits tighter filling head, short core, lower level of internal tip-burn,lower level of bottom rot, and the elevation of its base from theground.

The variety has shown uniformity and stability for the traits, withinthe limits of environmental influence for the traits. It has beenself-pollinated a sufficient number of generations with carefulattention to uniformity of plant type. The line has been increased withcontinued observation for uniformity. No variant traits have beenobserved or are expected in variety ‘E01E.70168’ Lot A.

The data which define these characteristics is based on observationstaken in San Juan Bautista, Calif. Color references are primarily to theRHS Colour Chart of The Royal Horticultural Society of London (RHS)(2007 edition). Descriptive terminology follows the Plant IdentificationTerminology, An Illustrated Glossary, 2nd edition by James G. Harris andMelinda Woolf Harris, unless where otherwise defined.

Objective Description of the Variety ‘E01E.70168’ Lot A

Lettuce variety ‘E01E.70168’ Lot A has the following morphologic andother characteristics:

Plant:

-   -   Type: Upright heading iceberg    -   Shape: Oblong to oval    -   Height: Tall    -   Diameter: Narrow    -   Core length: Relatively short under cool conditions; long cores        under high heat conditions

Leaves:

-   -   Heading:        -   Outer leaves: Curl over, less tight wrap than iceberg        -   Inner leaves: Blanched    -   Color:        -   Outer leaves: Medium green        -   Inner leaves: Yellow-green    -   Mature leaf texture: Crisp        Comparisons to Other Lettuce Varieties

Table 5 below compares the characteristics of lettuce variety‘E01E.70168’ Lot A with the most similar varieties ‘Pueblo’ (icebergtype), ‘True Heart’ (romaine type), and ‘Crunchita’ (cosberg type).Table 6 below compares the qualities of the harvested material of thesevarieties, including shelf life, maturity index, and other qualities.

TABLE 5 ‘E01E.70168’ ‘True Characteristic Lot A ‘Pueblo’ Heart’‘Crunchita’ Plant shape Oval to oblong Round V-shaped/ Oval to Deepoblong V-shaped Leaf shape Fan-shaped to Fan- V-shaped/ Fan-shapedV-shaped/Deep shaped Deep to V-shaped/ V-shaped (more V-shaped DeepFan-shaped) V-shaped (more V-shaped)

TABLE 6 ‘E01E.70168’ ‘True Characteristic Lot A ‘Pueblo’ Heart’‘Crunchita’ Shelf life (days post 18 22 16 16 harvest) Odor (days postharvest) 18 22 14 14 Browning (days post 18 22 16 16 harvest) Rotting(days post 16 20 14 14 harvest) Maturity index (1 to 9)  3.0 ± 0.5  4.1± 0.7 38.8 ± 17.7 36.8 ± 12.6 Processing efficiency (%) 93.2 ± 2.3 91.6± 4.1 84.7 ± 3.3  89.7 ± 2.7  Internal tipburn (1 to 9) 6.8 7.3 7.8 8.6Bremia (1 to 9) 7.9 7.7 9.0 9.0 Splitting (1 to 9) 4.5 4.3 6.0 8.6 Sideshoot (1 to 9) 8.6 9.0 9.0 9.0Overview of the Variety ‘E01E.70168’ Lot B

‘E01E.70168’ Lot B is an inbred line from an original cross between twoiceberg rogues. ‘E01E.70111‘ and’E01E.70168’ are inbred lines from thesame original cross. ‘E01E.70168‘ Lot A and’E01E.70168’ Lot B are sisterlines from the same parent in the previous generation.

Lettuce variety ‘E01E.70168’ Lot B is the result of numerous generationsof plant selections chosen for its oblong to oval shape, taller height,crisp leaf texture, and closed head. Further traits of interest includedits tighter filling head, short core, lower level of internal tip-burn,lower level of bottom rot, and the elevation of its base from theground.

The variety has shown uniformity and stability for the traits, withinthe limits of environmental influence for the traits. It has beenself-pollinated a sufficient number of generations with carefulattention to uniformity of plant type. The line has been increased withcontinued observation for uniformity. No variant traits have beenobserved or are expected in variety ‘E01E.70168’ Lot B.

The data which define these characteristics is based on observationstaken in San Juan Bautista, Calif. Color references are primarily to theRHS Colour Chart of The Royal Horticultural Society of London (RHS)(2007 edition). Descriptive terminology follows the Plant IdentificationTerminology, An Illustrated Glossary, 2nd edition by James G. Harris andMelinda Woolf Harris, unless where otherwise defined.

Objective Description of the Variety ‘E01E.70168’ Lot B

Lettuce variety ‘E01E.70168’ Lot B has the following morphologic andother characteristics:

Plant:

-   -   Type: Upright heading iceberg    -   Shape: Oblong to oval    -   Height: Tall    -   Diameter: Narrow    -   Core length: Relatively short under cool conditions; long cores        under high heat conditions

Leaves:

-   -   Heading:        -   Outer leaves: Curl over, less tight wrap than iceberg        -   Inner leaves: Blanched    -   Color:        -   Outer leaves: Medium green        -   Inner leaves: Yellow-green    -   Mature leaf texture: Crisp        Comparisons to Other Lettuce Varieties

Table 7 below compares the characteristics of lettuce variety‘E01E.70168’ Lot B with the most similar varieties ‘Pueblo’ (icebergtype), ‘True Heart’ (romaine type), and ‘Crunchita’ (cosberg type).Table 8 below compares the qualities of the harvested material of thesevarieties, including shelf life, maturity index, and other qualities.

TABLE 7 ‘E01E.70168’ ‘True Characteristic Lot B ‘Pueblo’ Heart’‘Crunchita’ Plant shape Oval to oblong Round V- shaped/ Oval to Deepoblong V-shaped Leaf shape Fan-shaped to Fan- V-shaped/ Fan-shapedV-shaped/Deep shaped Deep to V-shaped/ V-shaped (more V-shaped DeepFan-shaped) V-shaped (more V-shaped)

TABLE 8 ‘E01E.70168’ ‘True Characteristic Lot B ‘Pueblo’ Heart’‘Crunchita’ Shelf life (days post 16 22 16 16 harvest) Odor (days postharvest) 16 22 14 14 Browning (days post 16 22 16 16 harvest) Rotting(days post 16 20 14 14 harvest) Maturity index (1 to 9) 3.4 ± 0.7 4.1 ±0.7 38.8 ± 17.7 36.8 ± 12.6 Processing efficiency (%) 93.4 ± 1.3  91.6 ±4.1  84.7 ± 3.3  89.7 ± 2.7  Internal tipburn (1 to 9) 6.8 7.3 7.8 8.6Bremia (1 to 9) 7.9 7.7 9.0 9.0 Splitting (1 to 9) 6.0 4.3 6.0 8.6 Sideshoot (1 to 9) 8.6 9.0 9.0 9.0

DEPOSIT INFORMATION

A deposit of the lettuce variety ‘E01E.70111’ Lot A is maintained byEnza Zaden USA, Inc., having an address at 7 Harris Place, Salinas,Calif. 93901, United States. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to a deposit ofat least 2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of lettuce variety ‘E01E.70111’ Lot A were depositedon Jan. 5, 2018 according to the Budapest Treaty in the NationalCollection of Industrial, Food and Marine Bacteria Ltd (NCIMB Ltd),Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA,United Kingdom. The deposit has been assigned NCIMB number 42957. Accessto this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

A deposit of the lettuce variety ‘E01E.70111’ Lot B is maintained byEnza Zaden USA, Inc., having an address at 7 Harris Place, Salinas,Calif. 93901, United States. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to a deposit ofat least 2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of lettuce variety ‘E01E.70111’ Lot B were depositedon Jan. 22, 2018 according to the Budapest Treaty in the NationalCollection of Industrial, Food and Marine Bacteria Ltd (NCIMB Ltd),Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA,United Kingdom. The deposit has been assigned NCIMB number 42962. Accessto this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

A deposit of the lettuce variety ‘E01E.70168’ Lot A is maintained byEnza Zaden USA, Inc., having an address at 7 Harris Place, Salinas,Calif. 93901, United States. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to a deposit ofat least 2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of lettuce variety ‘E01E.70168’ Lot A were depositedon Jan. 5, 2018 according to the Budapest Treaty in the NationalCollection of Industrial, Food and Marine Bacteria Ltd (NCIMB Ltd),Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA,United Kingdom. The deposit has been assigned NCIMB number 42958. Accessto this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

A deposit of the lettuce variety ‘E01E.70168’ Lot B is maintained byEnza Zaden USA, Inc., having an address at 7 Harris Place, Salinas,Calif. 93901, United States. Access to this deposit will be availableduring the pendency of this application to persons determined by theCommissioner of Patents and Trademarks to be entitled thereto under 37C.F.R. § 1.14 and 35 U.S.C. § 122. Upon allowance of any claims in thisapplication, all restrictions on the availability to the public of thevariety will be irrevocably removed by affording access to a deposit ofat least 2,500 seeds of the same variety with the National Collection ofIndustrial, Food and Marine Bacteria Ltd. (NCIMB Ltd), FergusonBuilding, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, UnitedKingdom.

At least 2500 seeds of lettuce variety ‘E01E.70168’ Lot B were depositedon Jan. 22, 2018 according to the Budapest Treaty in the NationalCollection of Industrial, Food and Marine Bacteria Ltd (NCIMB Ltd),Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA,United Kingdom. The deposit has been assigned NCIMB number 42963. Accessto this deposit will be available during the pendency of thisapplication to persons determined by the Commissioner of Patents andTrademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35 U.S.C. §122. Upon allowance of any claims in this application, all restrictionson the availability to the public of the variety will be irrevocablyremoved.

The deposit will be maintained in the NCIMB depository, which is apublic depository, for a period of at least 30 years, or at least 5years after the most recent request for a sample of the deposit, or forthe effective life of the patent, whichever is longer, and will bereplaced if a deposit becomes nonviable during that period.

ENUMERATED EMBODIMENTS

The following enumerated embodiments are representative of some aspectsof the invention.

1. An upright heading iceberg lettuce plant comprising a height todiameter ratio greater than or equal to about 1.0, a height of about 1.5to about 3 times a height of a standard iceberg lettuce plant, a crispleaf texture, and a closed head.

2. The plant of embodiment 1, wherein the iceberg lettuce plant furthercomprises one or more characteristics selected from the group consistingof: a short core, a low level of internal tipburn, a low level of bottomrot, an increased number of leaves, a fast fill rate of leaves in ahead, and a space between a base of a head and a top of a ground.3. The plant of embodiment 2, wherein said space is achieved by anincreased length of core or an increased number of frame leaves outsideof a processing material of a head.4. An iceberg lettuce seed, wherein the seed produces an iceberg lettuceplant with an upright stature phenotype.5. The seed of embodiment 4, wherein the upright stature phenotypecomprises a height to diameter ratio greater than or equal to about 1.0,a height of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.6. The seed of embodiment 5, wherein the iceberg lettuce plant furthercomprises one or more characteristics selected from the group consistingof: a short core, a low level of internal tipburn, a low level of bottomrot, an increased number of leaves, a fast fill rate of leaves in ahead, and a space between a base of a head and a top of a ground.7. An iceberg lettuce seed comprising an upright stature allele at locusA, wherein the seed produces an iceberg lettuce plant with an uprightstature phenotype.8. The seed of embodiment 7, wherein the upright stature phenotypecomprises a height to diameter ratio greater than or equal to about 1.0,a height of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.9. The seed of embodiment 8, wherein the iceberg lettuce plant furthercomprises one or more characteristics selected from the group consistingof: a short core, a low level of internal tipburn, a low level of bottomrot, an increased number of leaves, a fast fill rate of leaves in ahead, and a space between a base of a head and a top of a ground.10. An iceberg lettuce plant produced from a cross of a first iceberglettuce plant with a second iceberg lettuce plant, wherein the firsticeberg lettuce plant produces an upright heading iceberg lettuce plantwith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head, the secondiceberg lettuce plant produces an upright heading iceberg lettuce plantwith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head, and wherein theiceberg lettuce plant produced from the cross has a height to diameterratio greater than or equal to about 1.0, a height of about 1.5 to about3 times a height of a standard iceberg lettuce plant, a crisp leaftexture, and a closed head.11. The plant of embodiment 10, wherein the first and second iceberglettuce plants are ‘E01E.70111’ Lot A lettuce plants, a sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957.12. The plant of embodiment 10, wherein the first and second iceberglettuce plants are ‘E01E.70111’ Lot B lettuce plants, a sample of‘E01E.70111’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42962.13. The plant of embodiment 10, wherein the first and second iceberglettuce plants are ‘E01E.70168’ Lot A lettuce plants, a sample of‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958.14. The plant of embodiment 10, wherein the first and second iceberglettuce plants are ‘E01E.70168’ Lot B lettuce plants, a sample of‘E01E.70168’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42963.15. The plant of embodiment 10, wherein the first iceberg lettuce plantis an ‘E01E.70111’ Lot A lettuce plant, a sample of ‘E01E.70111’ Lot Alettuce seed having been deposited under NCIMB Accession Number 42957.16. The plant of embodiment 10, wherein the second iceberg lettuce plantis an ‘E01E.70168’ Lot A lettuce plant, a sample of ‘E01E.70168’ Lot Alettuce seed having been deposited under NCIMB Accession Number 42958.17. An iceberg lettuce plant produced from a cross of an ‘E01E.70111’Lot A plant with a second ‘E01E.70111’ Lot A plant, a sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957, wherein the iceberg lettuce plant produced fromthe cross has a height to diameter ratio greater than or equal to about1.0, a height of about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head.18. An iceberg lettuce plant produced from a cross of an ‘E01E.70111’Lot B plant with a second ‘E01E.70111’ Lot B plant, a sample of‘E01E.70111’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42962, wherein the iceberg lettuce plant produced fromthe cross has a height to diameter ratio greater than or equal to about1.0, a height of about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head.19. An iceberg lettuce plant produced from a cross of an ‘E01E.70168’Lot A plant with a second ‘E01E.70168’ Lot A plant, a sample of‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958, wherein the iceberg lettuce plant produced fromthe cross has a height to diameter ratio greater than or equal to about1.0, a height of about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head.20. An iceberg lettuce plant produced from a cross of an ‘E01E.70168’Lot B plant with a second ‘E01E.70168’ Lot B plant, a sample of‘E01E.70168’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42963, wherein the iceberg lettuce plant produced fromthe cross has a height to diameter ratio greater than or equal to about1.0, a height of about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head.21. An iceberg lettuce plant produced from a cross of an ‘E01E.70111’Lot A plant with a second ‘E01E.70168’ Lot A plant, a sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957, and a sample of ‘E01E.70168’ Lot A lettuce seedhaving been deposited under NCIMB Accession Number 42958, wherein theiceberg lettuce seed produced from the cross produces an iceberg lettucewith a height to diameter ratio greater than or equal to about 1.0, aheight of about 1.5 to about 3 times a height of a standard iceberglettuce plant, a crisp leaf texture, and a closed head.22. An iceberg lettuce seed from the plants of any one of embodiments 1to 21.23. The plant of any one of embodiments 1 to 21, wherein the iceberglettuce plant has a height to diameter ratio greater than or equal toabout 1.0, a height of about 1.5 to about 3 times a height of a standardiceberg lettuce plant, a crisp leaf texture, and a closed head.24. A plant part from the plant of embodiment 23.25. The plant part of embodiment 24, wherein said part is a head, aseed, a leaf, or a portion thereof.26. The plant part of embodiment 25, wherein said part is a head.27. An iceberg lettuce plant having all the physiological andmorphological characteristics of the iceberg lettuce plant of embodiment23.28. A plant part from the plant of embodiment 27.29. The plant part of embodiment 28 wherein said part is a head, a leaf,or a portion thereof.30. The plant part of embodiment 29, wherein said part is a head.31. A pollen grain or an ovule of the plant of embodiment 27.32. A tissue culture produced from the plant of embodiment 27.33. An iceberg lettuce plant regenerated from the tissue culture ofembodiment 33, wherein the plant has all of the morphological andphysiological characteristics of an iceberg lettuce plant produced bygrowing seed designated as ‘E01E.70111’ Lot A, representative sample of‘E01E.70111’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42957.34. An iceberg lettuce plant regenerated from the tissue culture ofembodiment 33, wherein the plant has all of the morphological andphysiological characteristics of an iceberg lettuce plant produced bygrowing seed designated as ‘E01E.70111’ Lot B, representative sample of‘E01E.70111’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42962.35. An iceberg lettuce plant regenerated from the tissue culture ofembodiment 33, wherein the plant has all of the morphological andphysiological characteristics of an iceberg lettuce plant produced bygrowing seed designated as ‘E01E.70168’ Lot A, representative sample of‘E01E.70168’ Lot A lettuce seed having been deposited under NCIMBAccession Number 42958.36. An iceberg lettuce plant regenerated from the tissue culture ofembodiment 33, wherein the plant has all of the morphological andphysiological characteristics of an iceberg lettuce plant produced bygrowing seed designated as ‘E01E.70168’ Lot B, representative sample of‘E01E.70168’ Lot B lettuce seed having been deposited under NCIMBAccession Number 42963.37. A method of making iceberg lettuce seeds, said method comprisingcrossing the plant of embodiment 23 with another iceberg lettuce plantand harvesting seed therefrom.38. A method of making iceberg lettuce seeds, said method comprisingcrossing the plant of embodiment 27 with another iceberg lettuce plantand harvesting seed therefrom.39. A method of selecting an iceberg lettuce plant with upright stature,the method comprising:

-   -   a) crossing a first iceberg lettuce plant with a height to        diameter ratio greater than or equal to about 1.0, a height of        about 1.5 to about 3 times a height of a standard iceberg        lettuce plant, a crisp leaf texture, and a closed head with a        second iceberg lettuce plant with a height to diameter ratio        greater than or equal to about 1.0, a height of about 1.5 to        about 3 times a height of a standard iceberg lettuce plant, a        crisp leaf texture, and a closed head;    -   b) selecting offspring with an improved set of characteristics        comprising a height to diameter ratio greater than or equal to        about 1.5, a height greater than or equal to about 2 to about 3        times a height of a standard iceberg lettuce plant, a crisp leaf        texture, and a more tightly closed head;    -   c) selfing or sibbing said offspring; d) repeating steps b)        and c) for multiple generations to produce inbred lines with the        improved set of characteristics.        40. The method of embodiment 39, wherein the characteristics        used for selection in step b) further comprise one or more        characteristics selected from the group consisting of: one or        more characteristics selected from the group consisting of: a        short core, a low level of internal tipburn, a low level of        bottom rot, an increased number of leaves, a fast fill rate of        leaves in a head, and a space between a base of a head and a top        of a ground.        41. The method of embodiment 40, wherein said space between the        base of the head and the top of the ground is achieved by an        increased length of core or an increased number of frame leaves        outside of a processing material of a head.        42. An iceberg lettuce (Lactuca sativa) plant comprising a        genetic determinant that leads to the iceberg lettuce plant        having an upright stature, which genetic determinant is as        comprised in a lettuce plant representative seed of which was        deposited with the NCIMB under accession numbers 42957, 42962,        42958, and 42963.        43. The plant of embodiment 42, wherein the genetic determinant        is homozygously present.        44. The plant of embodiment 42, wherein the genetic determinant        is heterozygously present.        45. The plant of embodiment 42, wherein the upright stature        comprises one or more characteristics selected from the group        consisting of: a ratio of plant height to diameter greater than        about 1, a height of about 1.5 to about 3 times a height of a        standard iceberg lettuce plant, a crisp leaf texture, and a        closed head.        46. The plant of embodiment 42, wherein the lettuce further        comprises one or more characteristics selected from the group        consisting of: a short core, a low level of internal tipburn, a        low level of bottom rot, an increased number of leaves, a fast        fill rate of leaves in a head, and a space between a base of a        head and a top of a ground.        47. An iceberg lettuce (Lactuca sativa) plant comprising a        genetic determinant that leads to the iceberg lettuce plant        having an upright stature, wherein the upright stature comprises        one or more characteristics selected from the group consisting        of: a ratio of plant height to diameter greater than about 1, a        height of about 1.5 to about 3 times a height of a standard        iceberg lettuce plant, a crisp leaf texture, and a closed head,        and wherein the genetic determinant is as comprised in a lettuce        plant representative seed of which was deposited with the NCIMB        under accession numbers 42957, 42962, 42958, and 42963.        48. The plant of embodiment 47, wherein the genetic determinant        is homozygously present.        49. The plant of embodiment 47, wherein the genetic determinant        is heterozygously present.        50. The plant of embodiment 47, wherein the lettuce further        comprises one or more characteristics selected from the group        consisting of: a short core, a low level of internal tipburn, a        low level of bottom rot, an increased number of leaves, a fast        fill rate of leaves in a head, and a space between a base of a        head and a top of a ground.        51. The plant of any one of embodiments 42 to 50, obtainable by        crossing a first iceberg lettuce plant with a second iceberg        lettuce plant, wherein at least one of the said plants comprises        the genetic determinant as comprised in a lettuce plant        representative seed of which was deposited with the NCIMB under        accession numbers 42957, 42962, 42958, and 42963, or a progeny        plant thereof carrying the genetic determinant, and selecting,        preferably in the F₂ generation, for plants having an upright        stature.        52. A seed of the plant of any one of embodiments 42 to 50,        wherein the seed comprises the genetic determinant as present in        seeds of which a representative sample was deposited with the        NCIMB under accession numbers 42957, 42962, 42958, and 42963.        53. A progeny plant of the plant as embodied in embodiment 51,        wherein the plant comprises the genetic determinant as present        in seeds of which a representative sample was deposited with the        NCIMB under accession numbers 42957, 42962, 42958, or 42963.        54. The progeny plant as embodied in embodiment 53, wherein the        genetic determinant is homozygously present.        55. The progeny plant as embodied in embodiment 53, wherein the        genetic determinant is heterozygously present.        56. A progeny plant of the seed as embodied in embodiment 52,        wherein the plant comprises the genetic determinant as present        in seeds of which a representative sample was deposited with the        NCIMB under accession numbers 42957, 42962, 42958, or 42963.        57. The progeny plant as embodied in embodiment 56, wherein the        genetic determinant is homozygously present.        58. The progeny plant as embodied in embodiment 56, wherein the        genetic determinant is heterozygously present.        59. A propagation material derived from the plant as embodied in        embodiment 51, wherein the plant comprises the genetic        determinant as present in seeds of which a representative sample        was deposited with the NCIMB under accession numbers 42957,        42962, 42958, and 42963.        60. A propagation material derived from the seed as embodied in        embodiment 52, wherein the plant comprises the genetic        determinant as present in seeds of which a representative sample        was deposited with the NCIMB under accession numbers 42957,        42962, 42958, and 42963.        61. A propagation material capable of growing into a plant as        embodied in embodiment 42.        62. A propagation material capable of growing into a plant as        embodied in embodiment 47.        63. The propagation material as embodied in embodiments 61 or        62, wherein the propagation material comprises a microspore,        pollen, ovary, ovule, embryo, embryo sac, egg cell, cutting,        root, root tip, hypocotyl, cotyledon, stem, leaf, flower,        anther, seed, meristem, protoplast, callus, or cell.        64. A tissue culture of the propagation material as embodied in        embodiments 61 or 62.        65. A plant part of the plant as embodied in embodiment 42,        wherein said part is a head, a leaf, or a portion thereof.        66. A plant part of the plant as embodied in embodiment 47,        wherein said part is a head, a leaf, or a portion thereof.        67. The plant part of embodiments 65 or 66, wherein said part is        a head.        68. A method for producing an iceberg lettuce plant comprising        an upright stature phenotype, the method comprising:    -   (a) crossing the lettuce plant of embodiment 42 with a second        iceberg lettuce plant, wherein said second iceberg lettuce plant        is a non-upright stature iceberg lettuce plant, to produce        progeny;    -   (b) using said progeny of (a) in a back-crossing breeding        program with the iceberg lettuce plant of embodiment 42 as the        recurrent parent for one or more generations, to produce a        progeny iceberg lettuce plant comprising the upright stature        phenotype.        69. A method for producing an iceberg lettuce plant comprising        an upright stature phenotype, the method comprising:    -   (a) crossing the iceberg lettuce plant of embodiment 47 with a        second iceberg lettuce plant, wherein said second iceberg        lettuce plant is a non-upright stature iceberg lettuce plant, to        produce progeny;    -   (b) using said progeny of (a) in a back-crossing breeding        program with the iceberg lettuce plant of embodiment 47 as the        recurrent parent for one or more generations, to produce a        progeny iceberg lettuce plant comprising the upright stature        phenotype.        70. The method of embodiments 68 or 69, wherein the upright        stature phenotype further comprises one or more characteristics        selected from the group consisting of: a ratio of plant height        to diameter greater than about 1, a height of about 1.5 to about        3 times a height of a standard iceberg lettuce plant, a crisp        leaf texture, a closed head, a short core, a low level of        internal tipburn, a low level of bottom rot, an increased number        of leaves, a fast fill rate of leaves in a head, and a space        between a base of a head and a top of a ground.        71. An upright heading iceberg lettuce plant comprising a space        between a base of a head and a top of a ground.        72. The plant of embodiment 71, wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem).        73. The plant of embodiment 72, wherein the upright heading        iceberg lettuce plant further comprises one or more        characteristics selected from the group consisting of: a height        to diameter ratio of about 1.3 to about 1.5, leaf strength        between about 300 to 400 grams, about 40 to 50% overlapping        leaves inside the processing material of the head, a short core        inside of the processing material of the head (i.e., internal        stem), leaves with about 4.8 to 5.3° Brix, leaves with about 25        to 40 g/l total sugars, leaves with about 4.5 to 5% dry matter        weight, 5-10% processing damage per bag of processed plant        material, and a range of about 35 to 42 days of shelf life of        processed and packaged plant material.        74. An iceberg lettuce seed, wherein the seed produces an        iceberg lettuce plant with an upright stature phenotype.        75. The seed of embodiment 74, wherein the upright stature        phenotype comprises a space between a base of a head and a top        of a ground.        76. The seed of embodiment 75, wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem).        77. The seed of embodiment 76, wherein the upright stature        phenotype further comprises one or more characteristics selected        from the group consisting of: a height to diameter ratio of        about 1.3 to about 1.5, leaf strength between about 300 to 400        grams, about 40 to 50% overlapping leaves inside the processing        material of the head, a short core inside of the processing        material of the head (i.e., internal stem), leaves with about        4.8 to 5.3° Brix, leaves with about 25 to 40 g/l total sugars,        leaves with about 4.5 to 5% dry matter weight, 5-10% processing        damage per bag of processed plant material, and a range of about        35 to 42 days of shelf life of processed and packaged plant        material.        78. An iceberg lettuce plant produced from a cross of a first        iceberg lettuce plant with a second iceberg lettuce plant,        wherein the first iceberg lettuce plant is an upright heading        iceberg lettuce plant comprising a space between a base of a        head and a top of a ground, wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem), the second iceberg lettuce plant is        an upright heading iceberg lettuce plant comprising a space        between a base of a head and a top of a ground, wherein said        space comprises an increased length of core outside of a        processing material of a head (i.e., external stem), and wherein        the iceberg lettuce plant produced from the cross is an upright        heading iceberg lettuce plant comprising a space between a base        of a head and a top of a ground, wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem).        79. The plant of embodiment 78, wherein the first and second        iceberg lettuce plants are ‘E01E.70111’ Lot A lettuce plants, a        sample of ‘E01E.70111’ Lot A lettuce seed having been deposited        under NCIMB Accession Number 42957.        80. The plant of embodiment 78, wherein the first and second        iceberg lettuce plants are ‘E01E.70111’ Lot B lettuce plants, a        sample of ‘E01E.70111’ Lot B lettuce seed having been deposited        under NCIMB Accession Number 42962.        81. The plant of embodiment 78, wherein the first and second        iceberg lettuce plants are ‘E01E.70168’ Lot A lettuce plants, a        sample of ‘E01E.70168’ Lot A lettuce seed having been deposited        under NCIMB Accession Number 42958.        82. The plant of embodiment 78, wherein the first and second        iceberg lettuce plants are ‘E01E.70168’ Lot B lettuce plants, a        sample of ‘E01E.70168’ Lot B lettuce seed having been deposited        under NCIMB Accession Number 42963.        83. The plant of embodiment 78, wherein the first iceberg        lettuce plant is an ‘E01E.70111’ Lot A lettuce plant, a sample        of ‘E01E.70111’ Lot A lettuce seed having been deposited under        NCIMB Accession Number 42957.        84. The plant of embodiment 78, wherein the second iceberg        lettuce plant is an ‘E01E.70168’ Lot A lettuce plant, a sample        of ‘E01E.70168’ Lot A lettuce seed having been deposited under        NCIMB Accession Number 42958.        85. An iceberg lettuce plant produced from a cross of an        ‘E01E.70111’ Lot A plant with a second ‘E01E.70111’ Lot A plant,        a sample of ‘E01E.70111’ Lot A lettuce seed having been        deposited under NCIMB Accession Number 42957, wherein the        iceberg lettuce plant produced from the cross comprises a space        between a base of a head and a top of a ground, and wherein said        space comprises an increased length of core outside of a        processing material of a head.        86. An iceberg lettuce plant produced from a cross of an        ‘E01E.70111’ Lot B plant with a second ‘E01E.70111’ Lot B plant,        a sample of ‘E01E.70111’ Lot B lettuce seed having been        deposited under NCIMB Accession Number 42962, wherein the        iceberg lettuce plant produced from the cross comprises a space        between a base of a head and a top of a ground, and wherein said        space comprises an increased length of core outside of a        processing material of a head (i.e., external stem).        87. An iceberg lettuce plant produced from a cross of an        ‘E01E.70168’ Lot A plant with a second ‘E01E.70168’ Lot A plant,        a sample of ‘E01E.70168’ Lot A lettuce seed having been        deposited under NCIMB Accession Number 42958, wherein the        iceberg lettuce plant produced from the cross comprises a space        between a base of a head and a top of a ground, and wherein said        space comprises an increased length of core outside of a        processing material of a head (i.e., external stem).        88. An iceberg lettuce plant produced from a cross of an        ‘E01E.70168’ Lot B plant with a second ‘E01E.70168’ Lot B plant,        a sample of ‘E01E.70168’ Lot B lettuce seed having been        deposited under NCIMB Accession Number 42963, wherein the        iceberg lettuce plant produced from the cross comprises a space        between a base of a head and a top of a ground, and wherein said        space comprises an increased length of core outside of a        processing material of a head (i.e., external stem).        89. An iceberg lettuce plant produced from a cross of an        ‘E01E.70111’ Lot A plant with a second ‘E01E.70168’ Lot A plant,        a sample of ‘E01E.70111’ Lot A lettuce seed having been        deposited under NCIMB Accession Number 42957, and a sample of        ‘E01E.70168’ Lot A lettuce seed having been deposited under        NCIMB Accession Number 42958, wherein the iceberg lettuce plant        produced from the cross comprises a space between a base of a        head and a top of a ground, and wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem).        90. An iceberg lettuce seed from the plants of any one of        embodiments 71 to 89.        91. The plant of any one of embodiments 71 to 89, wherein the        iceberg lettuce plant comprises a space between a base of a head        and a top of a ground, and wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem).        92. The plant of embodiment 91, wherein the iceberg lettuce        plant further comprises one or more characteristics selected        from the group consisting of: a height to diameter ratio of        about 1.3 to about 1.5, leaf strength between about 300 to 400        grams, about 40 to 50% overlapping leaves inside the processing        material of the head, a short core inside of the processing        material of the head (i.e., internal stem), leaves with about        4.8 to 5.3° Brix, leaves with about 25 to 40 g/l total sugars,        leaves with about 4.5 to 5% dry matter weight, 5-10% processing        damage per bag of processed plant material, and a range of about        35 to 42 days of shelf life of processed and packaged plant        material.        93. A plant part from the plant of embodiment 92.        94. The plant part of embodiment 93, wherein said part is a        head, a seed, a leaf, or a portion thereof.        95. The plant part of embodiment 94, wherein said part is a        head.        96. An iceberg lettuce plant having all the physiological and        morphological characteristics of the iceberg lettuce plant of        embodiment 92.        97. A plant part from the plant of embodiment 96.        98. The plant part of embodiment 97 wherein said part is a head,        a leaf, or a portion thereof.        99. The plant part of embodiment 98, wherein said part is a        head.        100. A pollen grain or an ovule of the plant of embodiment 96.        101. A tissue culture produced from the plant of embodiment 96.        102. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 101, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70111’ Lot A, representative sample of ‘E01E.70111’ Lot A        lettuce seed having been deposited under NCIMB Accession Number        42957.        103. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 101, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70111’ Lot B, representative sample of ‘E01E.70111’ Lot B        lettuce seed having been deposited under NCIMB Accession Number        42962.        104. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 101, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70168’ Lot A, representative sample of ‘E01E.70168’ Lot A        lettuce seed having been deposited under NCIMB Accession Number        42958.        105. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 101, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70168’ Lot B, representative sample of ‘E01E.70168’ Lot B        lettuce seed having been deposited under NCIMB Accession Number        42963.        106. A method of making iceberg lettuce seeds, said method        comprising crossing the plant of embodiment 92 with another        iceberg lettuce plant and harvesting seed therefrom.        107. A method of making iceberg lettuce seeds, said method        comprising crossing the plant of embodiment 96 with another        iceberg lettuce plant and harvesting seed therefrom.        108. A method of making upright heading iceberg lettuce plants,        the method comprising:    -   a) crossing a first upright heading iceberg lettuce plant        comprising a space between a base of a head and a top of a        ground, wherein said space comprises an increased length of core        outside of a processing material of a head (i.e., external stem)        with a second upright heading iceberg lettuce plant comprising a        space between a base of a head and a top of a ground, wherein        said space comprises an increased length of core outside of a        processing material of a head (i.e., external stem);    -   b) selecting offspring lettuce plants comprising a space between        a base of a head and a top of a ground, wherein said space        comprises an increased length of core outside of a processing        material of a head (i.e., external stem);    -   c) selfing or sibbing said offspring;    -   d) repeating steps b) and c) for multiple generations to produce        inbred lines.        109. The method of embodiment 108, wherein the characteristics        used for selection in step b) further comprise one or more        characteristics selected from the group consisting of: a height        to diameter ratio of about 1.3 to about 1.5, leaf strength        between about 300 to 400 grams, about 40 to 50% overlapping        leaves inside the processing material of the head, a short core        inside of the processing material of the head (i.e., internal        stem), leaves with about 4.8 to 5.3° Brix, leaves with about 25        to 40 g/l total sugars, leaves with about 4.5 to 5% dry matter        weight, 5-10% processing damage per bag of processed plant        material, and a range of about 35 to 42 days of shelf life of        processed and packaged plant material.        110. An upright heading iceberg lettuce plant comprising a space        between a base of a head and a top of a ground.        111. The plant of embodiment 110, wherein said space comprises        an increased length of core outside of a processing material of        a head (i.e., external stem).        112. The plant of embodiment 111, wherein the increased length        of core outside of the processing material of the head (i.e.,        external stem) is about 3.5 cm or more, about 4 cm or more,        about 4.5 cm or more, or about 5 cm or more.        113. The plant of embodiment 111 or embodiment 112, wherein the        upright heading iceberg lettuce plant further comprises one or        more characteristics selected from the group consisting of: a        height to diameter ratio of about 1.3 to about 1.5, leaf        strength between about 300 to 400 grams, about 40 to 50%        overlapping leaves inside the processing material of the head, a        short core inside of the processing material of the head (i.e.,        internal stem), leaves with about 4.8 to 5.3° Brix, leaves with        about 25 to 40 g/l total sugars, leaves with about 4.5 to 5% dry        matter weight, 5-10% processing damage per bag of processed        plant material, and a range of about 35 to 42 days of shelf life        of processed and packaged plant material.        114. An upright heading iceberg lettuce seed, wherein the seed        produces an upright heading iceberg lettuce plant with an        upright stature phenotype.        115. The seed of embodiment 114, wherein the upright stature        phenotype comprises a space between a base of a head and a top        of a ground.        116. The seed of embodiment 115, wherein said space comprises an        increased length of core outside of a processing material of a        head (i.e., external stem).        117. The seed of embodiment 116, wherein the increased length of        core outside of the processing material of the head (i.e.,        external stem) is about 3.5 cm or more, about 4 cm or more,        about 4.5 cm or more, or about 5 cm or more.        118. The seed of embodiment 116 or embodiment 117, wherein the        upright stature phenotype further comprises one or more        characteristics selected from the group consisting of: a height        to diameter ratio of about 1.3 to about 1.5, leaf strength        between about 300 to 400 grams, about 40 to 50% overlapping        leaves inside the processing material of the head, a short core        inside of the processing material of the head (i.e., internal        stem), leaves with about 4.8 to 5.3° Brix, leaves with about 25        to 40 g/l total sugars, leaves with about 4.5 to 5% dry matter        weight, 5-10% processing damage per bag of processed plant        material, and a range of about 35 to 42 days of shelf life of        processed and packaged plant material.        119. An upright heading iceberg lettuce plant produced from a        cross of a first iceberg lettuce plant with a second iceberg        lettuce plant, wherein the first iceberg lettuce plant is an        upright heading iceberg lettuce plant comprising a space between        a base of a head and a top of a ground, wherein said space        comprises an increased length of core outside of a processing        material of a head (i.e., external stem), the second iceberg        lettuce plant is an upright heading iceberg lettuce plant        comprising a space between a base of a head and a top of a        ground, wherein said space comprises an increased length of core        outside of a processing material of a head (i.e., external        stem), and wherein the iceberg lettuce plant produced from the        cross is an upright heading iceberg lettuce plant comprising a        space between a base of a head and a top of a ground, wherein        said space comprises an increased length of core outside of a        processing material of a head (i.e., external stem).        120. The plant of embodiment 119, wherein the first and second        upright heading iceberg lettuce plants are ‘E01E.70111’ Lot A        lettuce plants, a sample of ‘E01E.70111’ Lot A lettuce seed        having been deposited under NCIMB Accession Number 42957.        121. The plant of embodiment 119, wherein the first and second        upright heading iceberg lettuce plants are ‘E01E.70111’ Lot B        lettuce plants, a sample of ‘E01E.70111’ Lot B lettuce seed        having been deposited under NCIMB Accession Number 42962.        122. The plant of embodiment 119, wherein the first and second        upright heading iceberg lettuce plants are ‘E01E.70168’ Lot A        lettuce plants, a sample of ‘E01E.70168’ Lot A lettuce seed        having been deposited under NCIMB Accession Number 42958.        123. The plant of embodiment 119, wherein the first and second        upright heading iceberg lettuce plants are ‘E01E.70168’ Lot B        lettuce plants, a sample of ‘E01E.70168’ Lot B lettuce seed        having been deposited under NCIMB Accession Number 42963.        124. The plant of embodiment 119, wherein the first upright        heading iceberg lettuce plant is an ‘E01E.70111’ Lot A lettuce        plant, a sample of ‘E01E.70111’ Lot A lettuce seed having been        deposited under NCIMB Accession Number 42957.        125. The plant of embodiment 78, wherein the second upright        heading iceberg lettuce plant is an ‘E01E.70168’ Lot A lettuce        plant, a sample of ‘E01E.70168’ Lot A lettuce seed having been        deposited under NCIMB Accession Number 42958.        126. An upright heading iceberg lettuce plant produced from a        cross of an ‘E01E.70111’ Lot A plant with a second ‘E01E.70111’        Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seed having        been deposited under NCIMB Accession Number 42957, wherein the        upright heading iceberg lettuce plant produced from the cross        comprises a space between a base of a head and a top of a        ground, and wherein said space comprises an increased length of        core outside of a processing material of a head.        127. An upright heading iceberg lettuce plant produced from a        cross of an ‘E01E.70111’ Lot B plant with a second ‘E01E.70111’        Lot B plant, a sample of ‘E01E.70111’ Lot B lettuce seed having        been deposited under NCIMB Accession Number 42962, wherein the        upright heading iceberg lettuce plant produced from the cross        comprises a space between a base of a head and a top of a        ground, and wherein said space comprises an increased length of        core outside of a processing material of a head (i.e., external        stem).        128. An upright heading iceberg lettuce plant produced from a        cross of an ‘E01E.70168’ Lot A plant with a second ‘E01E.70168’        Lot A plant, a sample of ‘E01E.70168’ Lot A lettuce seed having        been deposited under NCIMB Accession Number 42958, wherein the        upright heading iceberg lettuce plant produced from the cross        comprises a space between a base of a head and a top of a        ground, and wherein said space comprises an increased length of        core outside of a processing material of a head (i.e., external        stem).        129. An upright heading iceberg lettuce plant produced from a        cross of an ‘E01E.70168’ Lot B plant with a second ‘E01E.70168’        Lot B plant, a sample of ‘E01E.70168’ Lot B lettuce seed having        been deposited under NCIMB Accession Number 42963, wherein the        upright heading iceberg lettuce plant produced from the cross        comprises a space between a base of a head and a top of a        ground, and wherein said space comprises an increased length of        core outside of a processing material of a head (i.e., external        stem).        130. An upright heading iceberg lettuce plant produced from a        cross of an ‘E01E.70111’ Lot A plant with a second ‘E01E.70168’        Lot A plant, a sample of ‘E01E.70111’ Lot A lettuce seed having        been deposited under NCIMB Accession Number 42957, and a sample        of ‘E01E.70168’ Lot A lettuce seed having been deposited under        NCIMB Accession Number 42958, wherein the upright heading        iceberg lettuce plant produced from the cross comprises a space        between a base of a head and a top of a ground, and wherein said        space comprises an increased length of core outside of a        processing material of a head (i.e., external stem).        131. An upright heading iceberg lettuce seed from the upright        heading lettuce plants of any one of embodiments 110 to 130.        132. The upright heading iceberg lettuce plant of any one of        embodiments 110 to 130, wherein the iceberg lettuce plant        comprises a space between a base of a head and a top of a        ground, and wherein said space comprises an increased length of        core outside of a processing material of a head (i.e., external        stem).        133. The upright heading iceberg lettuce plant of any one of        embodiments 110 to 130, wherein the increased length of core        outside of the processing material of the head (i.e., external        stem) is about 3.5 cm or more, about 4 cm or more, about 4.5 cm        or more, or about 5 cm or more.        134. The plant of embodiment 132 or embodiment 133, wherein the        iceberg lettuce plant further comprises one or more        characteristics selected from the group consisting of: a height        to diameter ratio of about 1.3 to about 1.5, leaf strength        between about 300 to 400 grams, about 40 to 50% overlapping        leaves inside the processing material of the head, a short core        inside of the processing material of the head (i.e., internal        stem), leaves with about 4.8 to 5.3° Brix, leaves with about 25        to 40 g/l total sugars, leaves with about 4.5 to 5% dry matter        weight, 5-10% processing damage per bag of processed plant        material, and a range of about 35 to 42 days of shelf life of        processed and packaged plant material.        135. The plant of any one of embodiments 132 to 134, wherein the        external stem length allows for a harvest by machine.        136. The plant of embodiment 135, wherein the harvest by machine        does not remove a substantial portion of processing material of        a head.        137. The plant of embodiment 136, wherein the harvest by machine        is more efficient than for iceberg varieties ‘Reliant’ and        ‘Steamboat’, cosberg varieties ‘Cosmopolitan’ and ‘Crunchita’,        and romaine varieties ‘True Heart’ and ‘Solid King’.        138. A plant part from the plant of any one of embodiments 134        to 137.        139. The plant part of embodiment 138, wherein said part is a        head, a seed, a leaf, or a portion thereof.        140. The plant part of embodiment 139, wherein said part is a        head.        141. An iceberg lettuce plant having all the physiological and        morphological characteristics of the iceberg lettuce plant of        any one of embodiments 134 to 137.        142. A plant part from the plant of embodiment 141.        143. The plant part of embodiment 142 wherein said part is a        head, a leaf, or a portion thereof.        144. The plant part of embodiment 143, wherein said part is a        head.        145. A pollen grain or an ovule of the plant of embodiment 141.        146. A tissue culture produced from the plant of embodiment 141.        147. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 146, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70111’ Lot A, representative sample of ‘E01E.70111’ Lot A        lettuce seed having been deposited under NCIMB Accession Number        42957.        148. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 146, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70111’ Lot B, representative sample of ‘E01E.70111’ Lot B        lettuce seed having been deposited under NCIMB Accession Number        42962.        149. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 146, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70168’ Lot A, representative sample of ‘E01E.70168’ Lot A        lettuce seed having been deposited under NCIMB Accession Number        42958.        150. An iceberg lettuce plant regenerated from the tissue        culture of embodiment 146, wherein the plant has all of the        morphological and physiological characteristics of an iceberg        lettuce plant produced by growing seed designated as        ‘E01E.70168’ Lot B, representative sample of ‘E01E.70168’ Lot B        lettuce seed having been deposited under NCIMB Accession Number        42963.        151. A method of making iceberg lettuce seeds, said method        comprising crossing the plant of any one of embodiments 134 to        137 with another iceberg lettuce plant and harvesting seed        therefrom.        152. A method of making iceberg lettuce seeds, said method        comprising crossing the plant of embodiment 141 with another        iceberg lettuce plant and harvesting seed therefrom.        153. A method of making upright heading iceberg lettuce plants,        the method comprising:    -   a) crossing a first upright heading iceberg lettuce plant        comprising a space between a base of a head and a top of a        ground, wherein said space comprises an increased length of core        outside of a processing material of a head (i.e., external stem)        with a second upright heading iceberg lettuce plant comprising a        space between a base of a head and a top of a ground, wherein        said space comprises an increased length of core outside of a        processing material of a head (i.e., external stem);    -   b) selecting offspring lettuce plants comprising a space between        a base of a head and a top of a ground, wherein said space        comprises an increased length of core outside of a processing        material of a head (i.e., external stem);    -   c) selfing or sibbing said offspring;    -   d) repeating steps b) and c) for multiple generations to produce        inbred lines.        154. The method of embodiment 153, wherein the increased length        of core outside of the processing material of the head (i.e.,        external stem) is about 3.5 cm or more, about 4 cm or more,        about 4.5 cm or more, or about 5 cm or more.        155. The method of embodiment 153 or embodiment 154, wherein the        characteristics used for selection in step b) further comprise        one or more characteristics selected from the group consisting        of: a height to diameter ratio of about 1.3 to about 1.5, leaf        strength between about 300 to 400 grams, about 40 to 50%        overlapping leaves inside the processing material of the head, a        short core inside of the processing material of the head (i.e.,        internal stem), leaves with about 4.8 to 5.3° Brix, leaves with        about 25 to 40 g/l total sugars, leaves with about 4.5 to 5% dry        matter weight, 5-10% processing damage per bag of processed        plant material, and a range of about 35 to 42 days of shelf life        of processed and packaged plant material.

EXAMPLES

The present disclosure will be more fully understood by reference to thefollowing examples. They should not, however, be construed as limitingthe scope of the present disclosure. It is understood that the examplesand embodiments described herein are for illustrative purposes only andthat various modifications or changes in light thereof will be suggestedto persons skilled in the art and are to be included within the spiritand purview of this application and scope of the appended claims.

Example 1: Shelf Life Trial

The following example describes a trial comparing the shelf lifecapabilities of eight lettuce varieties of different types (e.g.,iceberg, romaine, and upright heading iceberg). ‘E01E.70111‘and’E01E.70168’ are inbred lines from the same original cross.‘E01E.70111‘ Lot A and’E01E.70111’ Lot B are sister lines from the sameparent in the previous generation. ‘E01E.70168‘ Lot A and’E01E.70168’Lot B are also sister lines from the same parent in the previousgeneration.

Materials and Methods

Lettuce Varieties:

The lettuce varieties and types used in the study are presented in Table9.

TABLE 9 Lettuce varieties and types tested in the shelf life trialLettuce Variety Lettuce Type True Heart Romaine Crunchita Cosberg PuebloIceberg Steamboat Iceberg E01E.70111 Lot A Upright heading icebergE01E.70111 Lot B Upright heading iceberg E01E.70168 Lot A Uprightheading iceberg E01E.70168 Lot B Upright heading iceberg

Procedure:

This was a small plot trial, and plants were planted by individuals withsimple push planters using raw seed. The lettuce plants were grown inSan Juan Bautista, Calif., USA, and harvested on Jul. 21, 2017. Theharvested material was kept at 41° F. for two days before processing.Plastic bags (30PA 200*300 unperforated) were used to store theharvested material (135 g plant material per bag) under MAP conditions(e.g., cold storage). Beginning on day 6 after harvest, the appearanceand smell of the harvested material in the bags were evaluated every 2days. In addition, other qualities of the harvested material wereevaluated, including processing efficiency, internal tipburn, andsplitting. A minimum of 2 bags per variety was opened at each evaluationpoint. Evaluation of the harvested material continued until the materialwas fully decayed.

Results

The results of the appearance and smell evaluation are presented belowin Table 10. The number under the heading “Shelf Life” was the number ofdays from harvest time to visually not acceptable. The number under theheading “Odor” was the number of days from harvest time to bad odor. Thenumber under the heading “Browning” was the number of days from harvesttime to first browning. The number under the heading “Rotting” was thenumber of days from harvest time to first significant rotting symptoms.

TABLE 10 Appearance and smell evaluation Shelf Life Odor (days BrowningRotting (days post post (days post (days Lettuce Variety harvest)harvest) harvest) post harvest) True Heart 16 14 16 14 Crunchita 16 1416 14 Pueblo 22 22 22 20 Steamboat 32 30 32 28 E01E.70111 Lot A 22 22 2220 E01E.70111 Lot B 20 20 18 18 E01E.70168 Lot A 18 18 18 16 E01E.70168Lot B 16 16 16 16

As can be seen in Table 10, the romaine and cosberg varieties ‘TrueHeart’ and ‘Crunchita’ developed bad odor and first significant rottingsymptoms two weeks after harvest. These varieties were also visuallyunacceptable and developed first browning symptoms within 16 days afterharvest. Similarly, both lines of ‘E01E.70168’ (Lot A and Lot B) beganexhibiting significant rotting symptoms at 16 days after harvest, andbegan browning, developed bad odor, and became visually unacceptable at16 to 18 days after harvest.

In contrast, the iceberg variety ‘Pueblo’ began exhibiting thesecharacteristics 20 to 22 days after harvest. The iceberg variety‘Steamboat’ (U.S. Pat. No. 7,977,536) exhibited first significantrotting symptoms 28 days after harvest, and became visuallyunacceptable, began browning, and developed bad odor 2 to 4 days afterthat (at 30 to 32 days after harvest). Similarly, both lines of‘E01E.70111’ (Lot A and Lot B) began exhibiting significant rottingsymptoms at 18 to 20 days after harvest, and began browning, developedbad odor, and became visually unacceptable at 18 to 22 days afterharvest. ‘E01E.70111’ Lot A developed these qualities 2 to 4 days after‘E01E.70111’ Lot B (consistently at 22 days after harvest, in comparisonto 18 or 20 days after harvest).

FIG. 2 illustrates the visual quality scores of each of the lettucevarieties over the evaluation period. The variety ‘True Heart’ (darkblue line) rapidly declined in visual quality from days 8 to 10, and thevarieties ‘Crunchita’ (pink line) as well as both ‘E01E.70168’ lines(Lot A and Lot B, red and orange lines respectively) declined at asimilarly rapid rate. ‘E01E.70111’ Lot B (purple line) also had a rapiddecrease in visual quality during storage. In contrast, ‘Pueblo’ and‘E01E.70111’ Lot A (teal and light blue lines respectively) maintainedgood visual quality. ‘Steamboat’ (green line) maintained good visualquality for the longest time of any of the varieties, and in fact showedlittle decrease in visual quality for a span of 10 days (day 10 to day20 from harvest time).

FIG. 3 illustrates the intensity of browning of each of the lettucevarieties. At the top of FIG. 3, a visual representation of the ratingsis provided. A rating of 1 (see top of FIG. 3, right-most image) wasgiven when the veins were completely brown. A rating of 9 (see top ofFIG. 3, left-most image) was given when the veins were white and clean.As can be seen in the bar chart at the bottom of FIG. 3, ‘True Heart’,‘Crunchita’, and ‘Pueblo’ showed a higher intensity of browning than theother varieties. None of the varieties showed strong browning symptomsduring storage. In addition, the intensity of pinking was very low in‘E01E.70168’, ‘E01E.70111’, and ‘Steamboat’.

In FIG. 4, the intensity of rotting for each of the lettuce varieties isillustrated. A rating of 1 on the rotting scale indicated that theharvested material of the lettuce varieties was completely rotten, whilea rating of 9 indicated that no rotting was present. ‘E01E.70168’ (Lot Aand Lot B), ‘True Heart’, and ‘Crunchita’ showed more rotting intensitythan the other lettuce varieties at an early storage time. In contrast,‘E01E.70111’ (Lot A and Lot B), ‘Pueblo’, and ‘Steamboat’ had very lowrotting intensity.

FIG. 5 illustrates the leaf intactness for each of the lettucevarieties. A rating of 1 on the scale indicated completely damagedleaves, while a rating of 9 indicated intact leaves. ‘True Heart’appeared more damaged than other varieties during processing. The threevarieties that were significantly deteriorated during cold storage,namely ‘True Heart’, ‘Crunchita’, and ‘E01E.70168’, also had theshortest shelf life of the varieties in the study. Those with longershelf life, such as ‘E01E.70111’, ‘Pueblo’, and ‘Steamboat’, did notshow significant bruises after 10 days in storage.

In Table 11, the maturity index for each of the varieties is shown.

TABLE 11 Maturity index for each of the varieties Maturity Index LettuceStandard Variety Average Deviation True Heart 38.8 17.7 Crunchita 36.812.6 Pueblo 4.1 0.7 Steamboat 3.4 0.9 E01E.70111 A 2.4 0.5 E01E.70111 B1.8 0.7 E01E.70168 A 3.0 0.5 E01E.70168 B 3.4 0.7

FIG. 6 shows the stages of the maturity index for iceberg lettuce,wherein stages 3 and 4 are considered ideal for processing.

In Table 12, the results of the evaluation of other qualities of theharvested material are shown. The processing efficiency percentage wasthe percentage of harvested plant material viable for processing. Theremaining categories were all measured on a scale of 1 to 9. ForInternal tipburn and Bremia (i.e., infection with the downy mildewpathogen Bremia lactucae), a rating of 9 meant there was no tipburn orBremia, and a rating of 1 meant the harvested material was full oftipburn or Bremia. For splitting, a rating of 9 meant the harvestedmaterial was soft splitting and a rating of 1 meant it was hardsplitting. Finally, for side shoots, a rating of 9 meant the harvestedmaterial had no side shoots and a rating of 1 meant the harvestedmaterial was full of side shoots.

TABLE 12 Evaluation of other qualities of the harvested materialProcessing Efficiency (%) Lettuce Standard Internal Split- Side VarietyAverage Deviation Tipburn Bremia ting Shoot True Heart 84.7 3.3 7.8 9.06.0 9.0 Crunchita 89.7 2.7 8.6 9.0 8.6 9.0 Pueblo 91.6 4.1 7.3 7.7 4.39.0 Steamboat 94.5 1.1 7.8 7.8 6.0 9.0 E01E.70111 A 82.3 2.1 8.3 9.0 7.59.0 E01E.70111 B 81.1 2.2 6.8 9.0 7.1 9.0 E01E.70168 A 93.2 2.3 6.8 7.94.5 8.6 E01E.70168 B 93.4 1.3 6.8 7.9 6.0 8.6

As can be seen in Table 12, the processing efficiency of the romaine andcosberg varieties ‘True Heart’ and ‘Crunchita’ was lower (84.7% and89.7%, respectively) than that of the iceberg varieties ‘Pueblo’ and‘Steamboat’ (91.6% and 94.5%, respectively). The processing efficiencyof both lines of ‘E01E.70111’ (Lot A and Lot B) was lower than either‘True Heart’ or ‘Crunchita’ (82.3% and 81.1%). In contrast, theprocessing efficiency of both lines of ‘E01E.70168’ (Lot A and Lot B)was intermediate between the two iceberg varieties (93.2% and 93.4%).

The cosberg variety ‘Crunchita’ had the lowest level of internal tipburn(rated 8.6 out of 9, where 9 is no internal tipburn), and ‘E01E.70111’Lot A had the second-lowest level (rated 8.3 out of 9). The romainevariety ‘True Heart’, as well as the two iceberg varieties ‘Pueblo’ and‘Steamboat’, had intermediate levels of internal tipburn (rated 7.8,7.3, and 7.8 out of 9, respectively). The varieties ‘E01E.70111’ Lot B,‘E01.70168’ A, and ‘E01.70168’ B had the highest levels of internaltipburn (rated 6.8 out of 9).

‘True Heart’, ‘Crunchita’, and both Lots of ‘E01E.70111’ had nomeasurable infection with the downy mildew pathogen B. lactucae. Incontrast, both iceberg varieties and both Lots of ‘E01E.70168’ hadmeasurable B. lactucae infection.

Of the varieties, the iceberg variety ‘Pueblo’ was the variety with themost hard splitting (rated 4.3 out of 9, where 9 was soft splitting and1 was hard splitting), and ‘E01E.70168’ Lot A was the variety with thesecond-most hard splitting (rated 4.5 out of 9). The cosberg variety‘Crunchita’ was the variety with the most soft splitting (rated 8.6 outof 9). The remaining varieties had intermediate splitting levels/types,with ratings ranging from 6.0 to 7.5.

Most of the lettuce varieties evaluated had no side shoots. Both linesof ‘E01E.70168’ (Lot A and Lot B), however, had a small number of sideshoots (rated 8.6 out of 9, where 9 was no side shoot and 1 was full ofside shoots).

Example 2: Trial Including Chop Test

This was a small plot trial to compare ‘E01E.70111‘ Lot Aand’E01E.70111’ Lot B characteristics. In addition, the harvestedmaterial was compared to a standard iceberg variety in a chop test. Thechop test was designed to test the qualities of chopped processingmaterial/product (i.e. a measurement of “chunking” or how easily thechopped sections of processing material/product separate). Product thatseparated easily when chopped would also mix more easily in a bag (e.g.,of prepared salad), while product that did not separate easily (i.e. thechopped sections stuck together) would mix less easily.

Materials and Methods

The plants were grown on 80 inch beds with 5 lines to a bed and thinnedto 10 inches between plants. Plants were planted by individuals withsimple push planters using raw seed.

Results

FIG. 7A illustrates the vertical cross-section of a head of lettucevariety ‘E01E.70111’ Lot B. From the cross-section, it can be seen thatthe head was cupped in, and the interior leaves were blanching, and hadrelatively short cores. There was fairly loose fill at this growingstage, and the plants were not yet completely mature.

In FIG. 7B, a side-by-side comparison of ‘E01E.70111‘ Lot Aand’E01E.70111’ Lot B is shown. The core of the Lot A head (on the left)appeared to be larger in diameter than the core of the Lot B head (onthe right), but these characteristics may not be generally applicable tothese lines. In general, larger diameter cores would help the plantstand up better in the field (more sturdy), but it could also mean thatthe core was longer which would not be as advantageous.

When an upright heading iceberg lettuce variety (e.g., ‘E01E.70111’ LotA) was compared to a standard iceberg lettuce variety in a chop test,the results were comparable (i.e., perhaps there was a slight colordifference) and could not be clearly distinguished by visual assessment.Therefore, it appeared that upright heading iceberg lettuce and standardiceberg lettuce had similar chunking characteristics.

Example 3: Genome-Wide Association Mapping

Molecular analysis will be performed on ‘E01E.70111’ Lot A, ‘E01E.70111’Lot B, ‘E01E.70168’ Lot A, and ‘E01E.70168’ Lot B. A genome-wideassociation mapping procedure will be used (e.g., GMAS, GWAS, etc.).This analysis aims to determine the genetic control of the physiologicaltype (i.e., upright heading iceberg lettuce).

Example 4: Size, Weight, Etc. Measurements

Large scale replicated trials will be planted. Plants will be seededwith a tractor and mechanical implements using pelleted seed. Plantingprocedure will be similar to commercial productions.

Measurements on head length, diameter, weight, maturity, and core lengthwill be taken. Additional measurements will also be taken, such ascalculated weight of harvested product per acre at different plantspacing.

Example 5: Large Scale Field Trial—Morphological Characterization

The following example describes a large scale trial comparing themorphological characteristics of ten lettuce varieties of differenttypes (e.g., cosberg, romaine, upright heading iceberg, and iceberg).The large scale trial included four separate tests conducted over thespring and summer of 2018 in California, USA.

Materials and Methods

Lettuce Varieties and Tests:

The lettuce varieties and types that were used in the study arepresented in Table 13. The four tests were conducted in Soledad, Calif.,USA, San Juan Bautista, Calif., USA, and Salinas, Calif., USA overspring and summer of 2018. The plant dates, harvest dates, days toharvest, season, and estimated trial maturity for the four tests in thelarge scale trial are presented in Table 14.

TABLE 13 Lettuce varieties and types tested in the large scale trialLettuce Variety Lettuce Type Cosmopolitan Cosberg Crunchita Cosberg TrueHeart Romaine Solid King Romaine E01E.70111 Lot A Upright headingiceberg E01E.70111 Lot B Upright heading iceberg E01E.70168 Lot AUpright heading iceberg E01E.70168 Lot B Upright heading iceberg ReliantIceberg Steamboat Iceberg

TABLE 14 Parameters of the four tests in the large scale trial SeasonEstimated Planting Harvest Days to California trial Test Date DateHarvest Central Coast maturity Location Test 1 Feb. May 18, 85 Spring85% Soledad, CA, USA 22, 2018 2018 Test 2 Feb. Jun. 5, 97 Spring 95% SanJuan Bautista, CA, 28, 2018 2018 USA Test 3 May 7, Jul. 10, 64 Summer80% Salinas, CA, USA 2018 2018 Test 4 May 8, Jul. 24, 77 Summer 100% SanJuan Bautista, CA, 2018 2018 USA

Growth and Harvesting:

All ten lettuce varieties were seeded, grown, and harvested in the sameway across all four tests. Plants were seeded with an EarthWay GardenSeeder (Model 1001-B) using pelleted (‘True Heart’, ‘Solid King’, and‘Reliant’) and raw seed (‘Cosmopolitan’, ‘Crunchita’, ‘E01E.70111’ LotA, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A, ‘E01E.70168’ Lot B, and‘Steamboat’). The harvest/postharvest procedures were identical acrossall ten lettuce varieties across all four tests. Lettuces were cutmanually by expert lettuce cutters at commercial maturity, and followingindustry standards. Lettuces were harvested that appeared healthy andwithout external damage in the field. Every block was harvested aroundthe same time of day, and all varieties were cut simultaneously toensure no variety remained in the sun for a much longer time than theother varieties. Twelve lettuce plants were harvested per variety pertest.

After harvest, lettuces were put immediately into boxes without removingthe external leaves. All boxes from one harvest were loaded in a randomorder onto the same truck for transport. The boxes were then transportedwithin two hours to a cold room kept at 41° F. (5° C.). For cold roomstorage, the harvested material was covered using plastic bags withperforations. Then, the material was stored for 48 hours beforeprocessing.

Morphological Characterization:

Six of the twelve plants were used for morphological characterization.At the beginning of each test, every variety received a random labnumber that was used for all evaluations in order to guarantee unbiasedassessments. On the day of processing, external leaves were carefullyremoved. Morphological evaluations were performed under natural lightand at room temperature (68° F.; 20° C.). For each test, evaluations ofall lettuce varieties were conducted in a random order over a period notlonger than 30 hours. In order to maintain the lettuces in optimalconditions between evaluations, lettuces were kept at 41° F. (5° C.),and then brought to room temperature. Data was taken by a postharvestresearcher and immediately transferred to a computer for use inpreparing analytical graphs and conducting statistical analyses.

Data Analysis:

Analytical graphs were prepared as box and whisker charts or bar charts.Box and whisker charts were used to show the distribution of the datainto quartiles, and to highlight the mean and outliers. The mean of thedata was shown by an “x” symbol within the box. Some of the boxes hadlines extending vertically (i.e., “whiskers”), which indicatedvariability outside the upper and lower quartiles. Any point that felloutside of the whiskers was considered an outlier, and was indicated bya dot symbol separate from both box and whiskers.

Bar charts were used to show the results of Analysis of Variance (ANOVA)statistical analyses. The bars in the bar charts depicted the means, andeach lettuce variety was shown as a differently-patterned bar (keys wereincluded at bottom of graphs; from left to right, bars corresponded tolettuce varieties ‘Cosmopolitan’, ‘Crunchita’, ‘True Heart’, ‘SolidKing’, ‘E01E.70111’ Lot A, ‘E01E.70111’ Lot B, ‘E01E.70168’ Lot A,‘E01E.70168’ Lot B, ‘Reliant’, and ‘Steamboat’). ANOVA was used todetermine whether there were any significant differences between themeans of independent (i.e., unrelated) groups, and the probability levelof alpha 5% was used in these analyses. If the difference between twomeans was found to be significant (i.e., the differences betweenlettuces from the same variety (variance within the same variety) weresmaller than the differences between varieties), the means were labeledwith different letters on the bar graph showing the results (letterswere shown at top of the bars on the graph). A result of a significantdifference showed that the compared varieties were different in regardto the characteristics of that trait. If the difference between twomeans was not found to be significant (i.e., the differences betweenlettuces from the same variety (variance within the same variety) weresimilar or higher than the differences between varieties), the meanswere labeled with the same letters on the bar graph showing the results(letters were shown at top of the bars on the graph). A result of nosignificant difference showed that the varieties were the same in regardto the characteristics of that trait. All ANOVAs were performed with thesoftware XLStat (Addinsoft 1995-2018, version 2018.5) with thespecifications: 2 interactions levels, 95% confidence interval, zeroconstraints, and pairwise comparison performed with Tukey test.

Measurement of Stem Length:

Stem length measurements were taken with a standard metric tape ininches. Measurements were later converted into centimeters (cm) forstatistical analyses. Two different stem length measurements were taken:external stem length and internal stem length. External stem wasmeasured from the harvest cut point at base of head to the first frameleaf of the head, and internal stem was measured from the first frameleaf of the head to the end of the stem at the center of the head(locations of external and internal stems are shown in FIG. 29). FIG.13A shows exemplary images of the external (left image) and internal(right image) stem length measurement procedures.

Measurement of Lettuce Head Weight:

Lettuce head weight measurements were taken with an Ohaus NV2101Precision Balance (2100 g capacity, 0.1 g readability). Weight wasmeasured in grams. An exemplary image of the lettuce head weightmeasurement procedure is shown in FIG. 14A.

Measurement of Lettuce Head Height and Width:

Lettuce head dimensions were taken with a standard metric tape ininches. Measurements were later converted into centimeters (cm) forstatistical analyses. Exemplary images of the lettuce head height (leftimage) and lettuce head width (right image) measurement procedures areshown in FIGS. 15A-15B.

Evaluation of the Percentage of Overlapping Leaves:

Evaluation of the percentage of overlapping leaves was done by visuallyevaluating individual lettuce heads. The lettuce heads were first cut inhalf, and then in quarters for the evaluation. All evaluations wereperformed by the same expert evaluator under direct light. An exemplaryimage of a lettuce head cut in half for use in the evaluation of thepercentage of overlapping leaves procedure is shown in FIG. 16A.

Measurement of Leaf Thickness:

Leaf thickness was measured with an electronic 0-1 inch digitalmicrometer caliper with 0.00005 inch/0.001 mm resolution (RexBeTi). Sixdata points were taken for each lettuce of each variety per test inorder to get a representative average of leaf thickness. Thesemeasurements were taken at the top and bottom of external, mid, andinternal leaves. Leaf thickness was measured in millimeters (mm).Exemplary images of the leaf thickness measurement procedure are shownin FIG. 17A.

Measurement of Leaf Color:

Two methods were used to measure the leaf color: Konica Minolta CM-700dSpectrophotometer (spectrophotometer) and the RHS Colour Chart of TheRoyal Horticultural Society of London, Sixth Edition 2015 (RHS ColourChart). The spectrophotometer was set to measure reflectance in theCIELAB color space (L*, a*, b*), 2 degree observer and C illuminant witha measurement area of 3 mm per measurement point (six measurements perlettuce, six lettuces per variety and test). The colorimeter was used inconjunction with SpectraMagic NX Professional software to recordmeasurements and to provide a comprehensive color analysis. Thespectrophotometer was used to measure three separate color components:lightness (i.e., brightness), hue angle (i.e., true color, arc tan(b*/a*, where b*=blue-yellow component and a*=green-red component))),and gloss (i.e., specular reflection at 8 degrees, shine). Exemplaryimages of the measurement of leaf color using the spectrophotometer areshown in FIG. 18A.

The RHS Colour Chart was used because it is the standard reference usedby horticulturists worldwide for recording plant colors. It has 920colors, and each color has a unique number and letter code as well as aname. An exemplary image of the measurement of leaf color using the RHScolour chart is shown in FIG. 19.

For both color measurement methods, six data points were taken for eachlettuce of each variety per test in order to get a representativeaverage of leaf color. These measurements were taken at the top andbottom of external, mid, and internal leaves.

Measurement of Leaf Strength:

Leaf strength was measured with the texture analyzer (texturometer)TA.XT Plus, which can measure forces from several hundredths of a gramup to 50 kilograms. In addition, the texturometer can travel at a speedranging from 0.01 and 40 mm/second, has micron level distanceresolution, and can be used with multiple probes and fixtures. For themeasurement of leaf strength (force (g) needed to tear a leaf or leaffragment), the TA-108s Single Small Film Extensibility Rig was attached.The procedure was set at a pre-test speed of 0.2 cm/sec, test speed of0.1 cm/sec, post-test speed of 1 cm/sec, a distance of 1.5 cm and atrigger force of 5 g. Six data points were taken for each lettuce ofeach variety per test in order to get a representative average of leafstrength. These measurements were taken at the top and bottom ofexternal, mid, and internal leaves. Exemplary images of the measurementof leaf strength using the texturometer are shown in FIGS. 20A-20B.

Results

Appearance of Lettuce Varieties:

Whole head and whole head vertical cross section images were taken foreach of the ten lettuce varieties in all four tests of the large scaletrial. FIGS. 9A-9J show exemplary images from Test 1, FIGS. 10A-10J showexemplary images from Test 2, FIGS. 11A-11J show exemplary images fromTest 3, and FIGS. 12A-12J show exemplary images from Test 4.

Measurement of Stem Length:

ANOVA analysis results of the means of external and internal stemlengths in cm from the four tests are shown in FIGS. 13B, 13E, 13H, 13K,and 13N. FIGS. 13C, 13F, 13I, 13L, and 13O show box and whisker chartsof external stem length data in cm from all four tests, and FIGS. 13D,13G, 13J, 13M, and 13P show box and whisker charts of internal stemlength data in cm from all four tests. Internal stem lengths of romaineand cosberg varieties were significantly longer than internal stemlengths of upright heading iceberg and iceberg varieties (FIG. 13N).Both upright heading iceberg and iceberg varieties had comparativelyshort internal stem lengths that were an average of 3-4 cm, whileromaine and cosberg varieties had long internal stem lengths of about5-6.25 cm. In regard to external stem lengths, upright heading icebergvarieties had an external stem that was significantly longer than theother tested varieties (FIG. 13N). The external stem length of uprightheading iceberg varieties was over 4 cm and up to almost 5 cm (for‘E01E.70111’ Lot A), while that of the other lettuce varieties was about2.5-3 cm.

Measurement of Lettuce Head Weight:

ANOVA analysis results of the means of lettuce head weight measurementin g from the four tests are shown in FIGS. 14B-14F. Head weights foreach variety varied across Tests 1-4. Overall, romaine variety ‘SolidKing’ consistently had the heaviest heads. The other varieties differedfrom test to test, for example in Test 3 (FIG. 14D) the icebergvarieties had the next-heaviest heads, while in Test 1 (FIG. 14B), theromaine and cosberg varieties had the next-heaviest heads along with theiceberg varieties and a couple of the upright heading iceberg lettucevarieties. When all tests were combined, the upright heading icebergvarieties were lighter in weight than the other varieties (FIG. 14F).

Measurement of Lettuce Head Height and Width:

ANOVA analysis results of the means of lettuce head height and widthmeasurements in cm from the four tests are shown in FIGS. 15B-15F.Overall, the heights of the cosberg, romaine, and machine harvestableiceberg lettuce varieties were comparable, while the iceberg lettucevarieties were significantly shorter in height (FIG. 15F). The widths ofall of the varieties were comparable, with the exception of the cosbergvariety ‘Cosmopolitan’ that was narrower than the rest of the varieties.

Evaluation of the Percentage of Overlapping Leaves:

ANOVA analysis results of the percentage of overlapping leaves from thefour tests are shown in FIGS. 16B-16F. In each of the individual tests,as well as when all tests were combined (FIG. 16F), the romainevarieties had a very low percentage of overlapping leaves at less than10%. The cosberg varieties had a slightly higher percentage ofoverlapping leaves, which was generally 10-20%. The upright headingiceberg varieties had an intermediate percentage of overlapping leaves,generally between 20-40%. Finally, the iceberg varieties had a very highpercentage of overlapping leaves at 80% or more.

Measurement of Leaf Thickness:

ANOVA analysis results of the means of leaf thickness measurements in cmfrom the four tests are shown in FIGS. 17B-17F. Leaf thicknesses werecomparable across all tested varieties in all tests (FIG. 17F).

Measurement of leaf color using the spectrophotometer: ANOVA analysisresults of the means of color lightness measurements from all four testsare shown in FIGS. 18B-18F. Color lightness was comparable betweenupright heading iceberg and iceberg varieties, and significantlydifferent from color lightness of romaine and cosberg varieties (FIG.18F). ANOVA analysis results of the means of color hue anglemeasurements from all four tests are shown in FIGS. 18G-18K. Color hueangle measurements were comparable across all of the varieties tested(FIG. 18K). ANOVA analysis results of the means of leaf glossmeasurements from all four tests is shown in FIGS. 18L-18P. As withcolor lightness measurements, leaf gloss was comparable between uprightheading iceberg and iceberg varieties, and significantly different fromleaf gloss of romaine and cosberg varieties (FIG. 18L).

Measurement of Leaf Color Using the RHS Colour Chart:

Six measurements were taken from each of the six tested plants for eachvariety per test, resulting in a total of 36 observations for eachvariety per test. The mode of each group of 36 observations was thenidentified in order to determine the representative color of thevariety. The mode colors of the ten tested varieties are listed in Table15 for Test 1, Table 16 for Test 2, Table 17 for Test 3, Table 18 forTest 4, and Table 19 for all tests (i.e., Test 1-4).

TABLE 15 Mode colors of all varieties in Test 1 Mode (RHS Mode VarietyObservations Categories value) frequency Color name Cosmopolitan 36 10143 7 Strong Yellow Green Crunchita 36 9 143 10 Strong Yellow Green TrueHeart 36 9 144 6 Strong Yellow Green Solid King 36 9 143 12 StrongYellow Green E01E.70111A 36 8 137 9 Moderate Olive Green E01E.70111B 369 141 8 Deep Yellowish Green E01E.70168A 36 8 145 12 Light yellow GreenE01E.70168B 36 7 144 10 Strong Yellow Green Reliant 36 10 145 7 LightYellow Green Steamboat 36 10 149 8 Brilliant Yellow Green

TABLE 16 Mode colors of all varieties in Test 2 Mode (RHS Mode VarietyObservations Categories value) frequency Color name Cosmopolitan 36 8144 11 Strong Yellow Green Crunchita 36 7 145 14 Light Yellow Green TrueHeart 36 10 144 8 Strong Yellow Green Solid King 36 7 143 9 StrongYellow Green E01E.70111A 36 7 145 13 Light Yellow Green E01E.70111B 36 9145 10 Light Yellow Green E01E.70168A 36 11 144 7 Strong Yellow GreenE01E.70168B 36 7 145 13 Light Yellow Green Reliant 36 9 145 12 LightYellow Green Steamboat 36 6 145 25 Light Yellow Green

TABLE 17 Mode colors of all varieties in Test 3 Mode (RHS Mode VarietyObservations Categories value) frequency Color name Cosmopolitan 36 6137 16 Moderate Olive Green Crunchita 36 7 144 12 Strong Yellow GreenTrue Heart 36 5 137 17 Moderate Olive Green Solid King 36 8 143 11Strong Yellow Green E01E.70111A 36 7 143 9 Strong Yellow GreenE01E.70111B 36 7 145 9 Light Yellow Green E01E.70168A 36 7 143 11 StrongYellow Green E01E.70168B 36 7 145 9 Light Yellow Green Reliant 36 8 14415 Strong Yellow Green Steamboat 36 7 145 22 Light Yellow Green

TABLE 18 Mode colors of all varieties in Test 4 Mode (RHS Mode VarietyObservations Categories value) frequency Color name Cosmopolitan 36 8137 11 Moderate Olive Green Crunchita 36 6 138 11 Moderate Olive GreenTrue Heart 36 7 137 12 Moderate Olive Green Solid King 36 7 143 14Strong Yellow Green E01E.70111A 36 6 145 17 Light Yellow GreenE01E.70111B 36 8 145 11 Light Yellow Green E01E.70168A 36 7 145 17 LightYellow Green E01E.70168B 36 7 144 11 Strong Yellow Green Reliant 36 6145 19 Light Yellow Green Steamboat 36 4 145 22 Light Yellow Green

TABLE 19 Mode colors of all varieties in all tests (i.e., Test 1-4) Mode(RHS Mode Variety Observations Categories value) frequency Color nameCosmopolitan 144 12 137 41 Moderate Olive Green Crunchita 144 12 145 31Light Yellow Green True Heart 144 11 137 39 Moderate Olive Green SolidKing 144 11 143 46 Strong Yellow Green E01E.70111A 144 9 145 38 LightYellow Green E01E.70111B 144 11 145 32 Light Yellow Green E01E.70168A144 12 145 41 Light Yellow Green E01E.70168B 144 11 144 35 Strong YellowGreen Reliant 144 13 145 46 Light Yellow Green Steamboat 144 11 145 75Light Yellow Green

These results showed that the iceberg varieties were clearlydistinguished from the romaine varieties by their color. Over all fourtests, the iceberg varieties were primarily evaluated to have the colorlight yellow green (RHS 145), while the romaine varieties were evaluatedto have either the color moderate olive green (RHS 137) or the colorstrong yellow green (RHS 143) (Table 19). The upright heading iceberglettuce varieties were evaluated to have the iceberg-type color of lightyellow green (RHS 145) across all four tests, with the exception of‘E01E.70168’ Lot B. Finally, the two cosberg varieties, ‘Cosmopolitan’and ‘Crunchita’, were together evaluated as having colors intermediatebetween romaine and iceberg over all four tests.

Measurement of Leaf Strength:

ANOVA analysis results of the means of leaf strength measurements in cmare shown in FIGS. 20C-20G. Overall, the leaf strength of uprightheading iceberg lettuce was comparable to the leaf strength of iceberglettuce (FIG. 20G). Both types of iceberg lettuce were significantlydifferent from romaine lettuce varieties, which had weaker leaves, anddifferent from cosberg lettuce varieties, which had stronger leaves.

Summary

In summary, the morphological characterization results showed thatupright heading iceberg varieties had an external stem that wassignificantly longer than the other tested varieties (i.e., romaine,cosberg, iceberg). These longer external stems raised the uprightheading iceberg variety heads off of the ground further than the headsof the other varieties, making these varieties suitable for mechanicalharvest. The mechanical harvest process uses uniform beds with levelground to ensure uniform cutting, and cutting occurs about 2 inchesabove the bed. This elevated cutting is done to prevent soil or otherdebris (e.g., leaf debris) from contaminating the product and beingtaken up into the cutting machine. Traditional iceberg lettuce varietieshave heads close to ground level, meaning that a large portion of thebottom of the head is cut off during mechanical harvest (FIG. 28). Thiscut both reduces yield and increases the risk of contaminating theproduct with other material (e.g., soil, leaf debris). Thus, the longerstem of the upright heading iceberg lettuce varieties situates the headat the correct height to be mechanically harvested without loss of headmaterial used for processing.

Moreover, the longer external stems of upright heading iceberg varietiesdid not result in longer internal stems. Longer internal stems are anegative characteristic for lettuce varieties used in processing,because when product is harvested and the stem is removed, there is lessproduct volume to use for processing. Further, the upright headingiceberg varieties had an intermediate percentage of overlapping leaves.The percentage of overlapping leaves is one way of assessing how cleanthe lettuce heads will be at harvest. The intermediate percentage ofoverlapping leaves of the upright heading iceberg varieties indicatesthat the heads of these varieties will be clean at harvest, similar tothe cosberg varieties, and in contrast to the romaine varieties.

In addition, upright heading iceberg varieties were shown to have manyof the same characteristics as iceberg varieties. These characteristicsincluded a short internal stem length and comparable head size. Further,the quality of the leaves was similar, with both leaf thickness and leafstrength showing no significant differences between upright headingiceberg and iceberg varieties. Finally, leaf appearance was shown to besimilar using multiple metrics. Overall, both the colorimeter(spectrophotometer) measurements and the RHS Colour Chart assessmentshowed that upright heading iceberg leaf color was comparable to icebergleaf color. Therefore, upright heading iceberg varieties had many of theknown and valued characteristics of iceberg varieties, while also havingthe new long external stem length characteristic that makes themsuitable for mechanical harvest.

Example 6: Large Scale Field Trial—Nutritional Characterization

The large scale trial described in Example 5 was also used to evaluatethe nutritional characteristics of the ten lettuce varieties ofdifferent types.

Materials and Methods

Lettuce Varieties and Tests:

Lettuce varieties and tests were as in Example 5.

Growth and Harvesting:

Growth and harvesting were as in Example 5.

Nutritional Characterization:

The same six of the twelve harvested plants (i.e., the same six plantsused for morphological characterization in Example 5) were evaluated fornutritional characteristics. At the beginning of each test, everyvariety received a random lab number that was used for all evaluationsin order to guarantee unbiased assessments. Morphological evaluationswere performed under natural light and at room temperature (68° F.; 20°C.). For each test, evaluations of all lettuce varieties were conductedin a random order over a period not longer than 30 hours. In order tomaintain the lettuces in optimal conditions between evaluations,lettuces were kept at 41° F. (5° C.), and then brought to roomtemperature. Data was taken by a postharvest researcher and immediatelytransferred to a computer for use in preparing analytical graphs andconducting statistical analyses.

Measurement of ° Brix:

° Brix measurement was done using the temperature-corrected MettlerToledo Refractometer 30GS (0.1% brix resolution). Around 300 g oflettuce plant material (mix of lettuce tissues including external, mid,and internal leaves, and upper and lower parts of each leaf type) wascut and mixed. Approximately 200 g of this mix of lettuce tissues werejuiced with a Nutribullet Pro 900 series (MagicBullet) and three dropsof this juice were immediately analyzed for ° brix. The average of three° brix measurements was considered the average ° brix of that lettuce.Six lettuces per variety and per test were analyzed in the study.

Measurement of Individual Sugars:

Around 300 g of lettuce plant material (mix of lettuce tissues includingexternal, mid, and internal leaves, and upper and lower parts of eachleaf type) was cut and mixed. Approximately 200 g of this mix of lettucetissues were juiced with a Nutribullet Pro 900 series (MagicBullet) and2 ml of juice was immediately analyzed for individual sugars using theSucrose/Fructose/Glucose Assay Kit from Megazyme (UV-method, withspectrophotometer and 1.5 ml cuvettes). The average of the threetechnical repetitions of sugar measurements was considered the averagesugars of that lettuce. Six lettuces per variety and per test wereanalyzed in the study. The sweetness factor formula was used tocalculate the sweetness factor: Sweetnessfactor=0.75*glucose+1*sucrose+1.75*fructose.

Measurement of Percentage of Dry Matter Weight:

Around 300 g of lettuce plant material (mix of lettuce tissues includingexternal, mid, and internal leaves, and upper and lower parts of eachleaf type) was cut and mixed. Approximately 80 g of this mix wereseparated from the rest and three batches of 25 g were accuratelyweighed and then immediately put in an oven at 55° C. After 48 hours (orthe point when final weight no longer changed), samples were weighedagain. The percentage of dry matter weight was calculated as the ratioof dry weight to fresh weight multiplied by 100. The average of thethree technical repetitions of dry matter measurements was consideredthe average dry matter weight of that lettuce. Six lettuces per varietyand per test were analyzed in the study.

Results

Measurement of ° Brix:

° Brix is generally used as an indicator of the sweetness of fruits andvegetables, as it is a simple method for measuring the percentage oftotal soluble solids concentration. A higher ° Brix value indicates ahigher amount of dissolved solids such as sucrose, glucose, andfructose, as well as vitamins, minerals, amino acids, proteins, hormonesand other nutrients in the vegetable. It is estimated that in a healthyfruit or vegetable, approximately 80% of the ° Brix is represented bythe natural sugars (i.e., sucrose, glucose, and fructose). ° Brixanalysis results for all ten tested varieties across all four tests areshown in FIG. 21. These results show that the romaine and cosbergvarieties had a lower ° Brix of about 4-4.5, while the iceberg varietieshad a range of ° Brix from about 4.5 (‘Reliant’) to over 6(‘Steamboat’). The upright heading iceberg varieties consistently had anintermediate ° Brix of about 5-5.5.

Measurement of Individual Sugars:

Although ° Brix provides a general overview, to evaluate the sweetnessof a product, it is necessary to study the individual sugars. Reducingsugars (glucose and fructose) are the major components of the solublesolids measured by ° Brix; sucrose is also present, but in very smallquantities. The sweetness factor (i.e., sweetness perceived by theconsumer) is dependent on the percentage of each of these three sugarsrelative to the total sugar content, since not all sugars are perceivedwith the same intensity. The sweetness of fructose is perceived earlierthan that of sucrose or glucose, and the taste sensation reaches a peak(higher than that of sucrose) and diminishes more quickly than that ofsucrose. Fructose can also enhance other flavors in the system. Thesweetness factor formula takes into account the different perceptions ofsweetness elicited by each of the three sugars.

FIG. 22A shows ANOVA analysis of the means of glucose, fructose, andsucrose measurements in g/l from all tests (i.e., Tests 1-4). Glucoseamounts were comparable between all of the varieties, with romaine andcosberg varieties having the lowest amount of glucose (except romainevariety ‘True Heart’). Fructose amounts were also comparable between allof the varieties, and again the romaine and cosberg varieties had thelowest amounts, as did one iceberg variety (‘Reliant’). Sucrose amountsdiffered more clearly between the different lettuce varieties: romaineand cosberg, as well as iceberg ‘Reliant’, had the lowest sucrose levelsof around 5 g/l, iceberg ‘Steamboat’ had the highest sucrose levels ofaround 20 g/l, and upright heading iceberg had intermediate sucroselevels around 10-15 g/l. Total sugar measurements, shown in FIG. 22B,were similarly distributed. Significant differences in amounts were seenbetween the romaine and cosberg varieties as well as iceberg ‘Reliant’(about 20-25 g/l), iceberg ‘Steamboat’ (about 40 g/l), and uprightheading iceberg (about 30-35 g/l). FIG. 22C shows that the calculatedsweetness factor was similarly distributed between these three groups.Again, the upright heading iceberg varieties were intermediate, and hada sweetness factor of about 40.

Measurement of Percentage of Dry Matter Weight:

While total solid measurements as described above comprise only solublesolids (e.g., sugars, organic acids, lipids, minerals and pigments;measured with ° Brix), percent dry matter content includes soluble andinsoluble solids (e.g., sugars, organic acids, lipids, minerals,pigments, proteins, cellulose, hemicellulose, pectins andpolysaccharides). FIG. 23 shows ANOVA analysis of the means ofpercentage of dry matter weight from all tests (i.e., Tests 1-4). Theseresults clearly show that the percentage of dry matter weight wassignificantly lower for the two iceberg varieties, at about 4%. Bothromaine and cosberg varieties had the highest percentages, at over5.25%. Upright heading iceberg varieties had intermediate percentages,at about 4.8-5%.

Summary

In summary, the nutritional characterization indicated that uprightheading iceberg varieties had intermediate sweetness levels. Theseintermediate sweetness levels were similar to the sweetness levels oficeberg varieties, which are valued by consumers for their sweetness,and higher than the sweetness levels of romaine and cosberg varieties.

Further, upright heading iceberg varieties had an intermediatepercentage of dry matter weight. Iceberg lettuce varieties tend to havelower dry matter weight values than romaine and cosberg varieties. Thiswas seen in the dry matter weight percentage results, which showedromaine and cosberg varieties to have significantly higher percentagesof dry matter weight than iceberg varieties. The intermediate percentageof dry matter weight of the upright heading iceberg varieties indicatesthat these varieties have a higher soluble and insoluble solids (sugars,organic acids, lipids, minerals, pigments, proteins, cellulose,hemicellulose, pectins and polysaccharides) than iceberg varieties.

Example 7: Large Scale Field Trial—Shelf Life Evaluations

The large scale trial described in Example 5 was also used to evaluatethe shelf life of the ten lettuce varieties of different types.

Materials and Methods

Lettuce Varieties and Tests:

Lettuce varieties and tests were as in Example 5.

Growth and Harvesting:

Lettuce varieties and tests were as in Example 5.

Shelf Life Evaluation:

The other six of the twelve harvested plants (i.e., not the same sixplants used for morphological and nutritional characterization inExamples 5 and 6) were used for shelf life evaluation. At the beginningof each test, every variety received a random lab number that was usedfor all evaluations in order to guarantee unbiased assessments. Plantswere processed as described below. Standard lettuce packaging bags wereused (30PA 200*300 unperforated) with modified atmosphere conditions(around 2% oxygen inside the sealed bag), and 135 g plant material perbag. Bags were evaluated (i.e., visual evaluations and aromaevaluations) by opening a minimum of two bags per lab number every twodays. The initial evaluation was done at day six after harvest andevaluation continued until the plant material was fully decayed.

Preparation of Plant Material for Shelf Life Evaluation:

Individual lettuces were processed with the Nemco 55650-1½″ Slice Easylettuce cutter. The cutter was cleaned and disinfected between varietiesto avoid mixing materials and/or spreading diseases. Exemplary images ofplant material processing are shown in FIGS. 24A-24B. Immediately aftercutting, plant material was disinfected and dried. First, the plantmaterial was washed with bleach for 30 seconds (50 ppm NaOCl diluted inwater at 41 F (5° C.); the pH of the disinfection water was between 6and 7 (lowered using HCl as needed)). Second, the plant material wasrinsed with water at 41° F. (5° C.) for 30 seconds. Third, the plantmaterial was collected in colanders to drain. Fourth, the plant materialwas centrifuged for 4 minutes in a Delfield SALD-1 Salad Dryer 20Gallons to dry. Exemplary images of plant material disinfection anddrying are shown in FIG. 24C.

After drying, the plant material was filled into standard lettuce bags(30PA 20429580 cm; suitable for modified atmospheric conditions). Plantmaterial of each variety, consisting of material from a mixture of thelettuce heads from the same variety, washed and centrifuged together,was weighed and packaged to produce a total of 25 bags containing 135 gof plant material each. An exemplary image of plant material weighingand packaging is shown in FIG. 24D. Modified atmospheric conditions(MAP) were used in the bags, because MAP is the standard market practice(large cut surfaces need low oxygen to prevent oxidation and highrespiring material needs air). MAP was achieved by sealing bags with aband sealer at 180-200° C. Next, a corner on the top of the bag was cutand used to vacuum the bags and then flush them with food grade nitrogenthree consecutive times to reduce the oxygen content within the bag toless than 2%. This process was performed with a three-way valve withvacuum hoses on each side: one to connect with the nitrogen cylinder(provided by Airgas), one to connect to the vacuum pump, and oneequipped with a hose connector or pipette tip to enter the bag. Thissetup was used to ensure that there was no leaking between the entranceof the bag and the tip. After this process, the bag was completelysealed again. Exemplary images of the process to achieve MAP and thesealed bags with MAP are shown in FIGS. 24E-24F.

Evaluation of Percentage of Processing Damage:

Evaluation of percentage of processing damage was based on visualquality evaluations. Blind tests were performed by the same expertevaluator in the same laboratory under the same light and at the sametime of the day. Evaluations were performed at the beginning of storage,usually between 4 and 12 days after processing, five bags per varietywere evaluated at a time. Processing damage was characterized as damageresulting directly from the plant material preparation process (i.e.,cutting, disinfecting, and drying the plant material). Data was reportedas percentage of physically damaged leaves in the bag, independent ofthe extent of the damage. An exemplary image of packaged plant materialin storage is shown in FIG. 24G.

Evaluation of Days of Shelf Life:

Evaluation of days of shelf life was based on overall visual qualityevaluations. Blind tests were performed by the same expert evaluator inthe same laboratory under the same light and at the same time of theday. Evaluation was done every other day (e.g., days 8, 10, 12, 14, 16,etc.), and five bags per variety were evaluated at a time. Days werecounted from the day of harvest to the day the lettuce bags were notmarketable anymore. Characteristics including overall visual quality,decay, browning, loss of texture, and loss of freshness (including aromadeterioration) were taken into account. If aroma was being evaluated,bags were opened and allowed to equilibrate at room temperature for15-30 minutes before evaluation. The end of shelf life was the day onwhich that individual bag of lettuce would no longer be accepted forconsumption, and this could be on the basis of any one of thesecharacteristics or a combination of one or more of thesecharacteristics. Each bag was individually evaluated and scored for daysof shelf life without information from previous days. At the end of theshelf life evaluation, after 50 days of storage, processed plantmaterial of each variety was photographed from two bags, one wasphotographed inside the packaging and the other was photographed afterplant material had been removed from the packaging (FIGS. 27A-27J).

Results

Evaluation of Percentage of Processing Damage:

FIGS. 25A-25D, and 25F show box and whisker charts of percentage ofprocessing damage data from all four tests. ANOVA analysis results ofthe means of percentage of processing damage are shown in FIG. 25E. FIG.25F, depicting the range of percentage of processing damage across Tests1-4, clearly shows that romaine and cosberg varieties were moresusceptible to processing damage, in some cases reaching values of over50%. In contrast, iceberg and upright heading iceberg varieties werebarely damaged during processing, and generally had 10% or less damage.In addition, iceberg and upright heading iceberg had a smaller range ofpercent processing damage across Tests 1-4. These differences in percentprocessing damage were statistically significant, and divided into threedistinct categories (FIG. 25E). The romaine varieties had the highestpercentage of processing damage, at about 37-42%. The cosberg varietieshad an intermediate percentage of processing damage, at about 20-22%.The iceberg and upright heading iceberg varieties had a low percentageof processing damage, ranging from 5-10%. Notably, iceberg and uprightheading iceberg formed one category; there was no clear differencebetween the two different iceberg types.

Evaluation of Days of Shelf Life:

FIGS. 26A-26D, and 26F show box and whisker charts of days of shelf lifedata from all four tests. ANOVA analysis results of the means of days ofshelf life data are shown in FIG. 26E. FIG. 26F, depicting the range ofdays of shelf life across Tests 1-4, clearly shows that romaine andcosberg varieties had shorter shelf lives than iceberg and uprightheading iceberg varieties. These differences in shelf life werestatistically significant, and divided into two distinct categories(FIG. 26E). The romaine and cosberg varieties had relatively short shelflives, with about 20-27 days passing before they were rated as no longeracceptable for consumption. In contrast, the iceberg and upright headingiceberg varieties had longer shelf lives of about 30-37 days. As withthe percentage of processing damage evaluation, iceberg and uprightheading iceberg formed one category. Clear differences in visualappearance were also seen at the end of the shelf life trial (i.e.,after 50 days of storage). FIGS. 27C-27D show that the romaine varieties(‘True Heart’ and ‘Solid King’) had browning, wilting, and liquiddevelopment in the bag. One of the cosberg varieties (‘Cosmopolitan’,FIG. 27A) had a similar appearance, while the other cosberg variety(‘Crunchita’, FIG. 27B) appeared more green and fresh. Some samples ofone of the iceberg varieties had clear browning on its leaves, but mostof the bags appeared green and fresh (‘Reliant’, FIG. 27I). Theremaining iceberg variety (‘ Steamboat’, FIG. 27J), as well as all ofthe upright heading iceberg varieties (FIGS. 27E-27H), had a green andfresh appearance.

Summary

In summary, the shelf life evaluation results showed that uprightheading iceberg varieties are suitable for industrial processing andpackaging. For one, the upright heading iceberg varieties had a lowpercentage of processing damage, which was comparable to the icebergvarieties. For another, the upright heading iceberg varieties had a longshelf life. Moreover, in this study, most of the samples of the uprightheading iceberg varieties maintained a green and fresh appearance evenafter 35 days of storage. These results indicate that upright headingiceberg varieties are sufficiently robust for industrial processing andpackaging. Moreover, they have a long shelf life, making these varietiessuitable for longer-term storage and distribution.

What is claimed is:
 1. An upright heading iceberg lettuce plantcomprising an external core length greater than 3.5 cm and a height todiameter ratio between about 1.3 and about 1.5, wherein the uprightheading iceberg lettuce plant is produced from a cross of a firstupright heading iceberg lettuce plant comprising an external core lengthgreater than 3.5 cm and a height to diameter ratio between about 1.3 andabout 1.5 with a second upright heading iceberg lettuce plant comprisingan external core length greater than 3.5 cm and a height to diameterratio between about 1.3 and about 1.5, and wherein the first and secondupright heading iceberg lettuce plants are grown from lettuce seeds asthose having been deposited with the NCIMB under accession numbers42957, 42962, 42958 and
 42963. 2. The upright heading iceberg lettuceplant of claim 1, further comprising leaf strength between about 300grams and about 400 grams.
 3. The upright heading iceberg lettuce plantof claim 2, further comprising between about 40% and about 50%overlapping leaves inside a processing material of a head.
 4. A head ofthe upright heading iceberg lettuce plant of claim
 1. 5. The uprightheading iceberg lettuce plant of claim 3, further comprising leaves withabout 4.8° to about 5.3° Brix.
 6. The upright heading iceberg lettuceplant of claim 5, further comprising leaves with about 25 g/l to about40 g/l total sugars.
 7. The upright heading iceberg lettuce plant ofclaim 6, further comprising leaves with about 4.5% to about 5% drymatter weight.
 8. The upright heading iceberg lettuce plant of claim 7,further comprising between 5% and 10% processing damage per bag ofprocessed plant material and a range of about 35 to 42 days of shelflife of processed and packaged plant material.
 9. A seed, tissue, orplant part of the upright heading iceberg lettuce plant of claim 1.